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  • Product Name
    Phospho-AKT1/2/3 (Ser473) Antibody
  • Catalog No.
    AF0016
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity:
    Human, Mouse, Rat
  • Prediction:
    Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%)
  • UniProt
  • Mol.Wt.
    60kDa
  • Concentration
    1mg/ml
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Product Information

Alternative Names:Expand▼

AKT 1; AKT; AKT1; AKT1_HUMAN; MGC99656; PKB; PKB-ALPHA; PRKBA; Protein Kinase B Alpha; Protein kinase B; Proto-oncogene c-Akt; RAC Alpha; RAC; RAC-alpha serine/threonine-protein kinase; RAC-PK-alpha; Akt2; AKT2_HUMAN; HIHGHH; murine thymoma viral (v-akt) homolog-2; PKB; PKB beta; PKBB; PKBBETA; PRKBB; Protein kinase Akt 2; Protein kinase Akt-2; Protein kinase B beta; rac protein kinase beta; RAC-BETA; RAC-beta serine/threonine-protein kinase; RAC-PK-beta; v akt murine thymoma viral oncogene homolog 2; Akt3; AKT3 kinase; AKT3_HUMAN; DKFZp434N0250; MPPH; PKB gamma; PKBG; PRKBG; Protein kinase Akt-3; Protein Kinase AKT3; Protein kinase B gamma; RAC gamma; RAC gamma serine/threonine protein kinase; RAC-gamma serine/threonine-protein kinase; RAC-PK-gamma; RACPK Gamma; Serine threonine protein kinase Akt 3; Serine threonine protein kinase Akt3; STK 2; STK-2; STK2; V akt murine thymoma viral oncogene homolog 3 (protein kinase B, gamma); V akt murine thymoma viral oncogene homolog 3; V akt murine thymoma viral oncogene homolog 3 protein kinase B gamma;

Applications:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000

Reactivity:

Human, Mouse, Rat

Predicted Reactivity:

Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%)

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

Specificity:

Phospho-Akt(Ser473) Antibody detects endogenous levels of Akt only when phosphorylated at Sersine 473.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 12 months from date of receipt.

Immunogen Information

Immunogen:

A synthesized peptide derived from human Akt around the phosphorylation site of Ser473.

Uniprot:



>>Visit The Human Protein Atlas

Gene id:

Molecular Weight:

Observed Mol.Wt.: 60kDa.
Predicted Mol.Wt.: 56kDa.

Subcellular Location:

Cytoplasm. Nucleus. Cell membrane. Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A. Phosphorylation on Tyr-176 by TNK2 results in its localization to the cell membrane where it is targeted for further phosphorylations on Thr-308 and Ser-473 leading to its activation and the activated form translocates to the nucleus.

Tissue Specificity:

Expressed in prostate cancer and levels increase from the normal to the malignant state (at protein level). Expressed in all human cell types so far analyzed. The Tyr-176 phosphorylated form shows a significant increase in expression in breast cancers during the progressive stages i.e. normal to hyperplasia (ADH), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC) and lymph node metastatic (LNMM) stages.

Description:

an AGC kinase that plays a critical role in controlling the balance between survival and AP0ptosis. Phosphorylated and activated by PDK1 in the PI3 kinase pathway. Mediates survival signals downstream of PI3 kinase and several growth factor receptors by phosphorylating AP0pototic proteins. First found in a mouse transforming retrovirus. Tumorigenic in a mouse lymphoma model and activated (by phospho-Akt staining) and/or overexpressed in a number of cancers including breast, prostate, lung, pancreatic, liver, ovarian and colorectal. Inhibitor: RX-0201. Substrates include tuberin, Bad, Forkhead transcription factors, caspase-9, and glycogen synthase kinase-3.

Sequence:
        10         20         30         40         50
MSDVAIVKEG WLHKRGEYIK TWRPRYFLLK NDGTFIGYKE RPQDVDQREA
60 70 80 90 100
PLNNFSVAQC QLMKTERPRP NTFIIRCLQW TTVIERTFHV ETPEEREEWT
110 120 130 140 150
TAIQTVADGL KKQEEEEMDF RSGSPSDNSG AEEMEVSLAK PKHRVTMNEF
160 170 180 190 200
EYLKLLGKGT FGKVILVKEK ATGRYYAMKI LKKEVIVAKD EVAHTLTENR
210 220 230 240 250
VLQNSRHPFL TALKYSFQTH DRLCFVMEYA NGGELFFHLS RERVFSEDRA
260 270 280 290 300
RFYGAEIVSA LDYLHSEKNV VYRDLKLENL MLDKDGHIKI TDFGLCKEGI
310 320 330 340 350
KDGATMKTFC GTPEYLAPEV LEDNDYGRAV DWWGLGVVMY EMMCGRLPFY
360 370 380 390 400
NQDHEKLFEL ILMEEIRFPR TLGPEAKSLL SGLLKKDPKQ RLGGGSEDAK
410 420 430 440 450
EIMQHRFFAG IVWQHVYEKK LSPPFKPQVT SETDTRYFDE EFTAQMITIT
460 470 480
PPDQDDSMEC VDSERRPHFP QFSYSASGTA

Background

Function:

AKT1 is one of 3 closely related serine/threonine-protein kinases (AKT1, AKT2 and AKT3) called the AKT kinase, and which regulate many processes including metabolism, proliferation, cell survival, growth and angiogenesis. This is mediated through serine and/or threonine phosphorylation of a range of downstream substrates. Over 100 substrate candidates have been reported so far, but for most of them, no isoform specificity has been reported. AKT is responsible of the regulation of glucose uptake by mediating insulin-induced translocation of the SLC2A4/GLUT4 glucose transporter to the cell surface. Phosphorylation of PTPN1 at 'Ser-50' negatively modulates its phosphatase activity preventing dephosphorylation of the insulin receptor and the attenuation of insulin signaling. Phosphorylation of TBC1D4 triggers the binding of this effector to inhibitory 14-3-3 proteins, which is required for insulin-stimulated glucose transport. AKT regulates also the storage of glucose in the form of glycogen by phosphorylating GSK3A at 'Ser-21' and GSK3B at 'Ser-9', resulting in inhibition of its kinase activity. Phosphorylation of GSK3 isoforms by AKT is also thought to be one mechanism by which cell proliferation is driven. AKT regulates also cell survival via the phosphorylation of MAP3K5 (apoptosis signal-related kinase). Phosphorylation of 'Ser-83' decreases MAP3K5 kinase activity stimulated by oxidative stress and thereby prevents apoptosis. AKT mediates insulin-stimulated protein synthesis by phosphorylating TSC2 at 'Ser-939' and 'Thr-1462', thereby activating mTORC1 signaling and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1. AKT is involved in the phosphorylation of members of the FOXO factors (Forkhead family of transcription factors), leading to binding of 14-3-3 proteins and cytoplasmic localization. In particular, FOXO1 is phosphorylated at 'Thr-24', 'Ser-256' and 'Ser-319'. FOXO3 and FOXO4 are phosphorylated on equivalent sites. AKT has an important role in the regulation of NF-kappa-B-dependent gene transcription and positively regulates the activity of CREB1 (cyclic AMP (cAMP)-response element binding protein). The phosphorylation of CREB1 induces the binding of accessory proteins that are necessary for the transcription of pro-survival genes such as BCL2 and MCL1. AKT phosphorylates 'Ser-454' on ATP citrate lyase (ACLY), thereby potentially regulating ACLY activity and fatty acid synthesis. Activates the 3B isoform of cyclic nucleotide phosphodiesterase (PDE3B) via phosphorylation of 'Ser-273', resulting in reduced cyclic AMP levels and inhibition of lipolysis. Phosphorylates PIKFYVE on 'Ser-318', which results in increased PI3P-5 activity. The Rho GTPase-activating protein DLC1 is another substrate and its phosphorylation is implicated in the regulation cell proliferation and cell growth. AKT plays a role as key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation. Signals downstream of phosphatidylinositol 3-kinase (PI3K) to mediate the effects of various growth factors such as platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin and insulin-like growth factor I (IGF-I). AKT mediates the antiapoptotic effects of IGF-I. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. May be involved in the regulation of the placental development. Phosphorylates STK4/MST1 at 'Thr-120' and 'Thr-387' leading to inhibition of its: kinase activity, nuclear translocation, autophosphorylation and ability to phosphorylate FOXO3. Phosphorylates STK3/MST2 at 'Thr-117' and 'Thr-384' leading to inhibition of its: cleavage, kinase activity, autophosphorylation at Thr-180, binding to RASSF1 and nuclear translocation. Phosphorylates SRPK2 and enhances its kinase activity towards SRSF2 and ACIN1 and promotes its nuclear translocation. Phosphorylates RAF1 at 'Ser-259' and negatively regulates its activity. Phosphorylation of BAD stimulates its pro-apoptotic activity. Phosphorylates KAT6A at 'Thr-369' and this phosphorylation inhibits the interaction of KAT6A with PML and negatively regulates its acetylation activity towards p53/TP53.

Post-translational Modifications:

O-GlcNAcylation at Thr-305 and Thr-312 inhibits activating phosphorylation at Thr-308 via disrupting the interaction between AKT1 and PDPK1. O-GlcNAcylation at Ser-473 also probably interferes with phosphorylation at this site.Phosphorylation on Thr-308, Ser-473 and Tyr-474 is required for full activity. Activated TNK2 phosphorylates it on Tyr-176 resulting in its binding to the anionic plasma membrane phospholipid PA. This phosphorylated form localizes to the cell membrane, where it is targeted by PDPK1 and PDPK2 for further phosphorylations on Thr-308 and Ser-473 leading to its activation. Ser-473 phosphorylation by mTORC2 favors Thr-308 phosphorylation by PDPK1. Phosphorylated at Thr-308 and Ser-473 by IKBKE and TBK1. Ser-473 phosphorylation is enhanced by interaction with AGAP2 isoform 2 (PIKE-A). Ser-473 phosphorylation is enhanced in focal cortical dysplasias with Taylor-type balloon cells. Ser-473 phosphorylation is enhanced by signaling through activated FLT3. Dephosphorylated at Thr-308 and Ser-473 by PP2A phosphatase. The phosphorylated form of PPP2R5B is required for bridging AKT1 with PP2A phosphatase. Ser-473 is dephosphorylated by CPPED1, leading to termination of signaling.Ubiquitinated via 'Lys-48'-linked polyubiquitination by ZNRF1, leading to its degradation by the proteasome (By similarity). Ubiquitinated; undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. TRAF6-induced 'Lys-63'-linked AKT1 ubiquitination is critical for phosphorylation and activation. When ubiquitinated, it translocates to the plasma membrane, where it becomes phosphorylated. When fully phosphorylated and translocated into the nucleus, undergoes 'Lys-48'-polyubiquitination catalyzed by TTC3, leading to its degradation by the proteasome. Also ubiquitinated by TRIM13 leading to its proteasomal degradation. Phosphorylated, undergoes 'Lys-48'-linked polyubiquitination preferentially at Lys-284 catalyzed by MUL1, leading to its proteasomal degradation.Acetylated on Lys-14 and Lys-20 by the histone acetyltransferases EP300 and KAT2B. Acetylation results in reduced phosphorylation and inhibition of activity. Deacetylated at Lys-14 and Lys-20 by SIRT1. SIRT1-mediated deacetylation relieves the inhibition.

Subcellular Location:

Nucleus;Plasma membrane;

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Subunit Structure:

Interacts with BTBD10 (By similarity). Interacts with KCTD20 (By similarity). Interacts (via the C-terminus) with CCDC88A (via its C-terminus). Interacts with GRB10; the interaction leads to GRB10 phosphorylation thus promoting YWHAE-binding (By similarity). Interacts with AGAP2 (isoform 2/PIKE-A); the interaction occurs in the presence of guanine nucleotides. Interacts with AKTIP. Interacts (via PH domain) with MTCP1, TCL1A AND TCL1B. Interacts with CDKN1B; the interaction phosphorylates CDKN1B promoting 14-3-3 binding and cell-cycle progression. Interacts with MAP3K5 and TRAF6. Interacts with BAD, PPP2R5B, STK3 and STK4. Interacts (via PH domain) with SIRT1. Interacts with SRPK2 in a phosphorylation-dependent manner. Interacts with RAF1. Interacts with TRIM13; the interaction ubiquitinates AKT1 leading to its proteasomal degradation. Interacts with TNK2 and CLK2. Interacts (via the C-terminus) with THEM4 (via its C-terminus). Interacts with and phosphorylated by PDPK1. Interacts with PA2G4 (By similarity). Interacts with KIF14; the interaction is detected in the plasma membrane upon INS stimulation and promotes AKT1 phosphorylation (PubMed:24784001). Interacts with FAM83B; activates the PI3K/AKT signaling cascade (PubMed:23676467). Interacts with WDFY2 (via WD repeats 1-3) (PubMed:16792529). Forms a complex with WDFY2 and FOXO1 (By similarity). Interacts with FAM168A (PubMed:23251525). Interacts with SYAP1 (via phosphorylated form and BSD domain); this interaction is enhanced in a mTORC2-mediated manner in response to epidermal growth factor (EGF) stimulation and activates AKT1 (PubMed:23300339).

Similarity:

Binding of the PH domain to phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3) following phosphatidylinositol 3-kinase alpha (PIK3CA) activity results in its targeting to the plasma membrane. The PH domain mediates interaction with TNK2 and Tyr-176 is also essential for this interaction.The AGC-kinase C-terminal mediates interaction with THEM4.Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. RAC subfamily.

Research Fields

Research Fields:

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.(View pathway)
· Cellular Processes > Cell growth and death > Apoptosis.(View pathway)
· Cellular Processes > Transport and catabolism > Autophagy - animal.(View pathway)
· Cellular Processes > Cellular community - eukaryotes > Signaling pathways regulating pluripotency of stem cells.(View pathway)
· Cellular Processes > Cell growth and death > Cellular senescence.(View pathway)
· Environmental Information Processing > Signal transduction > TNF signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > MAPK signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > ErbB signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > FoxO signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > mTOR signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Apelin signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Ras signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > AMPK signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Rap1 signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Phospholipase D signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Jak-STAT signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > cAMP signaling pathway.(View pathway)
· Human Diseases > Cancers: Specific types > Pancreatic cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Renal cell carcinoma.(View pathway)
· Human Diseases > Cancers: Overview > Pathways in cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Gastric cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Glioma.(View pathway)
· Human Diseases > Cancers: Specific types > Colorectal cancer.(View pathway)
· Human Diseases > Cancers: Overview > Proteoglycans in cancer.
· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.(View pathway)
· Human Diseases > Cancers: Specific types > Acute myeloid leukemia.(View pathway)
· Human Diseases > Cancers: Specific types > Endometrial cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Breast cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia.(View pathway)
· Human Diseases > Cancers: Specific types > Small cell lung cancer.(View pathway)
· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.
· Human Diseases > Cancers: Overview > Central carbon metabolism in cancer.(View pathway)
· Human Diseases > Cancers: Overview > Choline metabolism in cancer.(View pathway)
· Human Diseases > Infectious diseases: Viral > Hepatitis C.
· Human Diseases > Infectious diseases: Viral > Measles.
· Human Diseases > Infectious diseases: Viral > Hepatitis B.
· Human Diseases > Cancers: Specific types > Melanoma.(View pathway)
· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.
· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).
· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.
· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.
· Human Diseases > Infectious diseases: Viral > Influenza A.
· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.
· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.
· Human Diseases > Infectious diseases: Viral > HTLV-I infection.
· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).
· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.
· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.
· Human Diseases > Cancers: Specific types > Prostate cancer.(View pathway)
· Organismal Systems > Immune system > Toll-like receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > Platelet activation.(View pathway)
· Organismal Systems > Immune system > T cell receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > Fc epsilon RI signaling pathway.(View pathway)
· Organismal Systems > Immune system > Fc gamma R-mediated phagocytosis.(View pathway)
· Organismal Systems > Endocrine system > Progesterone-mediated oocyte maturation.
· Organismal Systems > Endocrine system > Relaxin signaling pathway.
· Organismal Systems > Digestive system > Carbohydrate digestion and absorption.
· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.(View pathway)
· Organismal Systems > Circulatory system > Adrenergic signaling in cardiomyocytes.(View pathway)
· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.
· Organismal Systems > Endocrine system > Insulin signaling pathway.(View pathway)
· Organismal Systems > Endocrine system > Regulation of lipolysis in adipocytes.
· Organismal Systems > Endocrine system > Prolactin signaling pathway.(View pathway)
· Organismal Systems > Nervous system > Neurotrophin signaling pathway.(View pathway)
· Organismal Systems > Endocrine system > Glucagon signaling pathway.
· Organismal Systems > Nervous system > Dopaminergic synapse.
· Organismal Systems > Development > Osteoclast differentiation.(View pathway)
· Organismal Systems > Aging > Longevity regulating pathway.(View pathway)
· Organismal Systems > Nervous system > Cholinergic synapse.
· Organismal Systems > Aging > Longevity regulating pathway - multiple species.(View pathway)
· Organismal Systems > Immune system > Chemokine signaling pathway.(View pathway)
· Organismal Systems > Immune system > B cell receptor signaling pathway.(View pathway)
· Organismal Systems > Endocrine system > Estrogen signaling pathway.(View pathway)

Western blot analysis of extracts from UV treated 293/3T3/Hela, using Phospho-Akt(Ser473) Antibody. -/+ means absence or presence of N peptide(non-phospho peptide) and P peptide(phospho peptide)
AF0016 at 1/200 staining human lung cancer tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
AF0016 at 1/200 staining human lung cancer tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
Phospho-Akt(Ser473) Antibody for IHC in human brain tissue
AF0016 staining 293 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25°C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37°C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.
AF0016 staining 293 cells by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25°C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37°C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) antibody(Red), diluted at 1/600, was used as secondary antibody.
ELISA analysis of AF0016 showing specificity to Phospho-AKT1/2/3 (Ser473) peptide. Peptides concentration: 1ug/ml.
P-peptide: phospho-peptide; N-peptide: non-phospho-peptide.

Reference Citations:

1). Ma J et al. Inhibition of Bcl-2 Synergistically Enhances the Antileukemic Activity of Midostaurin and Gilteritinib in Preclinical Models of FLT3-mutated Acute Myeloid Leukemia. Clin Cancer Res 2019 Jul 18 (PubMed: 31320594) [IF=8.911]

2). Li X et al. Antileukemic activity and mechanism of action of the novel PI3K and histone deacetylase dual inhibitor CUDC-907 in acute myeloid leukemia. Haematologica 2019 Feb 28 (PubMed: 30819918) [IF=7.570]

3). Liu Y et al. Increased autophagy in EOC re-ascites cells can inhibit cell death and promote drug resistance. Cell Death Dis 2018 Apr 1;9(4):419 (PubMed: 29549251) [IF=5.959]

4). Duan J et al. DINP aggravates autoimmune thyroid disease through activation of the Akt/mTOR pathway and suppression of autophagy in Wistar rats. Environ Pollut 2018 Oct 30;245:316-324 (PubMed: 30447474) [IF=5.714]

5). Huang Y et al. LncRNA AK023391 promotes tumorigenesis and invasion of gastric cancer through activation of the PI3K/Akt signaling pathway. J Exp Clin Cancer Res 2017 Dec 28;36(1):194 (PubMed: 29282102) [IF=5.646]

6). Su Y et al. Targeting PI3K, mTOR, ERK, and Bcl-2 signaling network shows superior antileukemic activity against AML ex vivo. Biochem Pharmacol 2018 Feb;148:13-26 (PubMed: 29208365) [IF=4.825]

7). Wang J et al. High temperature suppressed SSC self-renewal through S phase cell cycle arrest but not apoptosis. Stem Cell Res Ther 2019 Jul 29;10(1):227 (PubMed: 31358059) [IF=4.627]

8). Li QZ et al. Immunomodulatory activity of Ganoderma lucidum immunomodulatory protein via PI3K/Akt and MAPK signaling pathways in RAW264.7 cells. J Cell Physiol 2019 May 30 (PubMed: 31148200) [IF=4.522]

9). Wei W et al. Inhibition of insulin resistance by PGE1 via autophagy-dependent FGF21 pathway in diabetic nephropathy. Sci Rep 2018 Jan 8;8(1):9 (PubMed: 29311680) [IF=4.011]

10). Guo Z et al. MAP kinase-interacting serine/threonine kinase 2 promotes proliferation, metastasis, and predicts poor prognosis in non-small cell lung cancer. Sci Rep 2017 Sep 6;7(1):10612 (PubMed: 28878291) [IF=4.011]

Application: WB    Species:human;    Sample:Not available

(b) Western bolt shown that inhibited MEK, both decrease MNK2 expression and eIF4E phosphorylation. Inhibited AKT, decreased MNK2 and eIF4E, 4EBP1 phosphorylation.


11). Liu Y et al. Oligo-Porphyran Ameliorates Neurobehavioral Deficits in Parkinsonian Mice by Regulating the PI3K/Akt/Bcl-2 Pathway. Mar Drugs 2018 Mar 6;16(3) (PubMed: 29509717) [IF=3.772]

12). Feng X et al. CXCL5, the upregulated chemokine in patients with uterine cervix cancer, in vivo and in vitro contributes to oncogenic potential of Hela uterine cervix cancer cells. Biomed Pharmacother 2018 Nov;107:1496-1504 (PubMed: 30257367) [IF=3.743]

13). Shen F et al. CEMIP promotes ovarian cancer development and progression via the PI3K/AKT signaling pathway. Biomed Pharmacother 2019 Mar 26;114:108787 (PubMed: 30925458) [IF=3.743]

14). He WH et al. Estradiol promotes trophoblast viability and invasion by activating SGK1. Biomed Pharmacother 2019 Jun 13;117:109092 (PubMed: 31203134) [IF=3.743]

15). Liu HJ et al. A Novel Partial Agonist of Peroxisome Proliferator-Activated Receptor γ with Excellent Effect on Insulin Resistance and Type 2 Diabetes. J Pharmacol Exp Ther 2015 Jun;353(3):573-81 (PubMed: 25876909) [IF=3.615]

16). Yu G et al. The protective effect of low-energy shock wave on testicular ischemia-reperfusion injury is mediated by the PI3K/AKT/NRF2 pathway. Life Sci 2018 Oct 12 (PubMed: 30321543) [IF=3.448]

17). Yao J et al. Chrysin alleviates allergic inflammation and airway remodeling in a murine model of chronic asthma. Int Immunopharmacol 2016 Mar;32:24-31 (PubMed: 26780233) [IF=3.361]

18). Yao J et al. Chrysin alleviates allergic inflammation and airway remodeling in a murine model of chronic asthma. Int Immunopharmacol 2016 Mar;32:24-31 (PubMed: 26780233) [IF=3.361]

Application: WB    Species:mouse;    Sample:mouse

Fig. 9. Chrysin inhibits the phosphorylation of Akt and ERK1/2. The levels of Akt, p-Akt, ERK1/2 and p-ERK1/2 in protein extracts from lung tissues were analyzed with Western blot (A). The bar graph represents percent phosphorylation level compared with the control group (B and C). Densitometric data are expressed as mean ± SEM (#p b 0.05 vs. control group; *p b 0.05 vs. OVA group, n = 6).


19). Song Z et al. The protective role of the MKP-5-JNK/P38 pathway in glucolipotoxicity-induced islet β-cell dysfunction and apoptosis. Exp Cell Res 2019 Jun 13 (PubMed: 31202710) [IF=3.329]

20). He Y et al. Quercetin reverses experimental pulmonary arterial hypertension by modulating the TrkA pathway. Exp Cell Res 2015 Nov 15;339(1):122-34 (PubMed: 26476374) [IF=3.329]

Application: WB    Species:rat;    Sample:rat

Fig. 7. Effect of quercetin on TrkA/AKT pathway signaling in PASMCs. (A) The expression levels of p-TrkA/TrkA and p-AKT/AKT were measured by Western blotting in PASMCs in hypoxia and treated with increasing concentrations of quercetin for 24 h. (B) Results were quantified by densitometry analysis of the bands form (A) and then normalization to GAPDH protein. (C) Effects of NGF, GW441756, and quercetin on the expression of p-TrkA/TrkA and p-AKT/AKT in PASMCs in hypoxia. (D) Densitometric quantification of the bands were shown. (E) The effect of quercetin on PASMCs migration ability under the hypoxia condition treated with or without NGF (50 μg/L). (F) Quantification of the number of cells migrating through the polycarbonate membrane of average of 3 independent experiments. (G) Quantification of the Flow cytometry analysis results to confirm the effects of quercetin on PASMCs apoptosis and (H) cell cycle under the hypoxia condition treated with NGF (50 ug/L). *Po0.05, **Po0.01 compared with control; #Po0.05, ##Po0.01 compared with hypoxia and quercetin treated groups; &Po0.05, &&Po0.01 compared with NGF treated PASMCs.


21). Li S et al. Effect of CAPE-pNO2 against type 2 diabetes mellitus via the AMPK/GLUT4/ GSK3β/PPARα pathway in HFD/STZ-induced diabetic mice. Eur J Pharmacol 2019 Mar 15;853:1-10 (PubMed: 30885574) [IF=3.170]

22). Ge H et al. DcR3 induces proliferation, migration, invasion, and EMT in gastric cancer cells via the PI3K/AKT/GSK-3β/β-catenin signaling pathway. Onco Targets Ther 2018 Jul 19;11:4177-4187 (PubMed: 30050309) [IF=3.046]

23). Zhang L et al. Transdifferentiation effects and related mechanisms of nerve growth factor and internal limiting membrane on Müller cells. Exp Eye Res 2018 Dec 20;180:146-154 (PubMed: 30578789)

24). Liao ZL et al. Salidroside protects PC-12 cells against amyloid β-induced apoptosis by activation of the ERK1/2 and AKT signaling pathways. Int J Mol Med 2019 Feb 1 (PubMed: 30720058)

25). Gu X et al. Molecular interplay between microRNA-130a and PTEN in palmitic acid-mediated impaired function of endothelial progenitor cells: Effects of metformin. Int J Mol Med 2019 Mar 20 (PubMed: 30896786)

26). Yang M et al. Estrogen receptor β exhibited anti-tumor effects on osteosarcoma cells by regulating integrin, IAP, NF-kB/BCL-2 and PI3K/Akt signal pathway. J Bone Oncol 2017 Sep 28;9:15-20 (PubMed: 29071206)

Application: WB    Species:human;    Sample:Not available

Fig 5. The expression of p-p65, p-AKT and Bcl-2 (protein level) in U2-OS cells treated by LY294002 or/and Estrogen receptor β (ERβ) siRNA in the presence of 10-10 M E2.


27). Liu Y et al. Fucoxanthin Activates Apoptosis via Inhibition of PI3K/Akt/mTOR Pathway and Suppresses Invasion and Migration by Restriction of p38-MMP-2/9 Pathway in Human Glioblastoma Cells. Neurochem Res 2016 Oct;41(10):2728-2751 (PubMed: 27394418)

Application: WB    Species:human;    Sample:Not available

Fig. 4 Fucoxanthin promotes apoptosis via inhibition of PI3K/Akt/ mTOR signaling pathway in glioblastoma cells. a Cell lysates were electrophoresed and Akt, p-Akt, mTOR and p-mTOR proteins were detected by their respective specific antibodies in indicated concentrations


28). Huang YX et al. Ovostatin 2 knockdown significantly inhibits the growth, migration, and tumorigenicity of cutaneous malignant melanoma cells. PLoS One 2018 Apr 23;13(4):e0195610 (PubMed: 29684087)

29). Yang H et al. Neuronal protective effect of Songling Xuemaikang capsules alone and in combination with carbamazepine on epilepsy in kainic acid-kindled rats. Pharm Biol 2019 Dec;57(1):22-28 (PubMed: 30724642)

30). Zong W et al. Ginsenoside compound K attenuates cognitive deficits in vascular dementia rats by reducing the Aβ deposition. J Pharmacol Sci 2019 Feb 13 (PubMed: 30799178)

31). Wang W et al. Umbilical cord‑derived mesenchymal stem cells can inhibit the biological functions of melanoma A375 cells. Oncol Rep 2018 Jul;40(1):511-517 (PubMed: 29767256)

32). Geng S et al. P53 modulates hepatic insulin sensitivity through NF-κB and p38/ERK MAPK pathways. Biochem Biophys Res Commun 2018 Jan 15;495(3):2139-2144 (PubMed: 29258820)

33). Xie XC et al. Relaxin Attenuates Contrast-Induced Human Proximal Tubular Epithelial Cell Apoptosis by Activation of the PI3K/Akt Signaling Pathway In Vitro. Biomed Res Int 2017;2017:2869405 (PubMed: 28540295)

Application: WB    Species:human;    Sample:Not available

Figure 5: Effects of RLX on the expression of pAkt and apoptosis proteins


34). Li C et al. TREM2 inhibits inflammatory responses in mouse microglia by suppressing the PI3K/NF-κB signaling. Cell Biol Int 2018 Apr 16 (PubMed: 29663649)

35). Li C et al. TREM2 inhibits inflammatory responses in mouse microglia by suppressing the PI3K/NF-κB signaling. Cell Biol Int 2018 Apr 16 (PubMed: 29663649)

36). Li C et al. TREM2 inhibits inflammatory responses in mouse microglia by suppressing the PI3K/NF-κB signaling. Cell Biol Int 2018 Apr 16 (PubMed: 29663649)

37). Vijay V et al. Interleukin-8 blockade prevents activated endothelial cell mediated proliferation and chemoresistance of acute myeloid leukemia. Leuk Res 2019 Jul 3;84:106180 (PubMed: 31299413)

38). Wang F et al. Angelicin inhibits liver cancer growth in vitro and in vivo. Mol Med Rep 2017 Oct;16(4):5441-5449 (PubMed: 28849216)

Application: WB    Species:human;    Sample:Not available

Treatment with angelicin alters the protein expression levels of PI3K, p-Akt and total Akt in HepG2 and Huh-7 cells in vitro. (A) Representative blots demonstrating PI3K, p-Akt and total Akt protein expression levels in HepG2 and Huh-7 cells, following treatment with various concentrations of angelicin. (B) PI3K blots were semi-quantified using densitometry analysis of the protein bands and normalized to GAPDH. (C) p-Akt/Akt blots were semi-quantified using densitometry analysis of the protein bands. (D) Effects of LY294002 on angelicin-induced apoptosis. Cells were treated with the PI3K inhibitor LY294002 (3 mM) for 1 h prior to treatment with angelicin. The percentage of apoptotic cells following treatment with angelicin in the presence or absence of LY294002 was assessed using Annexin V-fluorescein isothiocyanate/propidium iodide staining and flow cytometry. Data are expressed as the mean ± standard deviation of 3 independent experiments.


39). Feng F et al. Louqin Zhisou Decoction Inhibits Mucus Hypersecretion for Acute Exacerbation of Chronic Obstructive Pulmonary Disease Rats by Suppressing EGFR-PI3K-AKT Signaling Pathway and Restoring Th17/Treg Balance. Evid Based Complement Alternat Med 2019 Jan 20;2019:6471815 (PubMed: 30800170)

40). Li J et al. The Aqueous Extract of Gynura divaricata (L.) DC. Improves Glucose and Lipid Metabolism and Ameliorates Type 2 Diabetes Mellitus. Evid Based Complement Alternat Med 2018 Jan 23;2018:8686297 (PubMed: 29599810)

41). Chai L et al. Biological functions of lung cancer cells are suppressed in co-culture with mesenchymal stem cells isolated from umbilical cord. Exp Ther Med 2018 Jan;15(1):1076-1080 (PubMed: 29399109)

42). Ning C et al. Targeting ERK enhances the cytotoxic effect of the novel PI3K and mTOR dual inhibitor VS-5584 in preclinical models of pancreatic cancer. Oncotarget 2017 Jul 4;8(27):44295-44311 (PubMed: 28574828)

Application: WB    Species:human;    Sample:Not available

Figure 5: BVD-523 synergizes with VS-5584 in AML cells. (A) BxPC-3, MIAPaCa-2, and CFPAC-1 cells were treated with vehicle control or variable concentrations of BVD-523 and VS-5584, alone or in combination, for 48 h. Viable cells were measured by MTT assays. Standard isobologram analyses of the antitumor interactions are shown. The IC50 values of each drug are plotted on the axes; the solid line represents the additive effect, while the points represent the concentrations of each drug resulting in 50% inhibition of proliferation. Points falling below the line indicate synergism whereas those above the line indicate antagonism. (B) HPAC cells were treated with vehicle control, VS, BVD-523 (BVD), or VS plus BVD for 48 h. Cells were fxed with 80% ice-cold ethanol and stained with PI for cell cycle analysis. The percentage of cells with sub-G1 DNA content are graphed as means of triplicates ± SEM from one representative experiment. ***indicates p < 0.001; combined treatment compared to individual treatments and vehicle control. Combination index (CI) values were calculated using CompuSyn software. (C) CI vs. Fa plot (combination index vs. fraction affected) for the cell death data is presented. (D) HPAC cells were treated with vehicle control or the indicated drugs for 48 h. Whole cell lysates were subjected to Western blotting and probed with the indicated antibody. The fold changes for the densitometry measurements, normalized to β-actin and then compared to no drug treatment control, are indicated.


43). Liu J et al. Irisin Enhances Doxorubicin-Induced Cell Apoptosis in Pancreatic Cancer by Inhibiting the PI3K/AKT/NF-κB Pathway. Med Sci Monit 2019 Aug 14;25:6085-6096 (PubMed: 31412018)

44). et al. miR-205-5p regulates epithelial-mesenchymal transition by targeting PTEN via PI3K/AKT signaling pathway in cisplatin-resistant nasopharyngeal carcinoma cells.

45). Yu Z et al. Intracavernosal Adeno-Associated Virus-Mediated S100A1 Gene Transfer Enhances Erectile Function in Diabetic Rats by Promoting Cavernous Angiogenesis via VEGF-A/VEGFR2 Signaling. J Sex Med 2019 Aug 1 (PubMed: 31378707)

46). WenjingZong et al. Ginsenoside compound K attenuates cognitive deficits in vascular dementia rats by reducing the Ab deposition. Journal of Pharmacological Sciences 2019

47). Hu J et al. Ursodeoxycholic acid ameliorates hepatic lipid metabolism in LO2 cells by regulating the AKT/mTOR/SREBP-1 signaling pathway. World J Gastroenterol 2019 Mar 28;25(12):1492-1501 (PubMed: 30948912)

48). Huikun Xu et al. Chemerin promotes the viability and migration of human placental microvascular endothelial cells and activates MAPK/AKT signaling . Int J Clin Exp Med 2018;11(2):721-727

49). et al. Low Long Noncoding RNA Growth Arrest-Specific Transcript 5 Expression in the Exosomes of Lung Cancer Cells Promotes Tumor Angiogenesis.

50). et al. Stem cell extracellular matrix-modified decellularized tendon slices facilitate the migration of bone marrow mesenchymal stem cells.

51). et al. Protective effects of macamides from Lepidium meyenii Walp. against corticosterone-induced neurotoxicity in PC12 cells†.

Review: 这个抗体wb的稀释比可以做到1/4000,效果非常好,条带单一。

Anonymous | 2017-10-16


Review: Staining of mouse embryonic stem cells. Staining is very strong so high dilution is needed at 1:1000 or even more for cell culture.

Anonymous | 2016-12-11


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Catalog Number :

AF0016-BP
(Blocking peptide available as AF0016-BP)

Price/Size :

$200/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Chicken
100%
Rabbit
100%
Pig
100%
Dog
100%
Sheep
100%
Horse
100%
Bovine
100%
Zebrafish
0%
Xenopus
0%
High similarity Medium similarity Low similarity No similarity
P31749/P31751/Q9Y243 as Substrate
Site PTM Type Enzyme
S2 Phosphorylation
K8 Ubiquitination
K14 Acetylation
K14 Methylation
K14 Ubiquitination
K20 Acetylation
K20 Ubiquitination
K30 Ubiquitination
T34 Phosphorylation O15530 (PDPK1) , Q05513 (PRKCZ)
K39 Ubiquitination
T65 Phosphorylation
T72 Phosphorylation P31749 (AKT1)
T87 Phosphorylation
T92 Phosphorylation
S122 Phosphorylation
S124 Phosphorylation
S126 O-Glycosylation
S126 Phosphorylation
S129 O-Glycosylation
S129 Phosphorylation P68400 (CSNK2A1)
S137 Phosphorylation
K140 Ubiquitination
T146 Phosphorylation
K154 Ubiquitination
K158 Ubiquitination
T172 Phosphorylation
Y176 Phosphorylation Q07912 (TNK2)
K189 Ubiquitination
T211 Phosphorylation
S216 Phosphorylation
C224 S-Nitrosylation
S246 Phosphorylation P31749 (AKT1)
S259 Phosphorylation
K268 Ubiquitination
K276 Sumoylation
K276 Ubiquitination
K284 Ubiquitination
K289 Ubiquitination
T291 Phosphorylation
C296 S-Nitrosylation
K297 Ubiquitination
K301 Sumoylation
K301 Ubiquitination
T305 O-Glycosylation
T305 Phosphorylation
T308 O-Glycosylation
T308 Phosphorylation P31749 (AKT1) , P17252 (PRKCA) , Q9UHD2 (TBK1) , P05771 (PRKCB) , Q15139 (PRKD1) , Q8N5S9 (CAMKK1) , Q14164 (IKBKE) , Q07912 (TNK2) , P78527 (PRKDC) , O15530 (PDPK1)
C310 S-Nitrosylation
T312 O-Glycosylation
T312 Phosphorylation P49840 (GSK3A)
Y315 Phosphorylation Q13882 (PTK6) , P12931 (SRC) , P07949 (RET)
Y326 Phosphorylation P12931 (SRC) , Q13882 (PTK6)
K377 Ubiquitination
S378 Phosphorylation
S396 Phosphorylation
K400 Ubiquitination
Y417 Phosphorylation
K420 Acetylation
K426 Acetylation
K426 Ubiquitination
T435 Phosphorylation
Y437 Phosphorylation
T443 Phosphorylation
T448 Phosphorylation
T450 Phosphorylation P42345 (MTOR)
S457 Phosphorylation
S473 O-Glycosylation
S473 Phosphorylation P05771 (PRKCB) , Q9UHD2 (TBK1) , P78527 (PRKDC) , Q9Y2I7 (PIKFYVE) , Q96KB5 (PBK) , Q53ET0 (CRTC2) , Q14164 (IKBKE) , P17252 (PRKCA) , Q5S007 (LRRK2) , P31749 (AKT1) , O15530 (PDPK1) , Q13418 (ILK) , Q15139 (PRKD1) , Q07912 (TNK2) , P42345 (MTOR) , P49137 (MAPKAPK2) , P28482 (MAPK1) , Q13153 (PAK1)
Y474 Phosphorylation
S475 Phosphorylation
S477 Phosphorylation
T479 Phosphorylation
Site PTM Type Enzyme
M1 Acetylation
K14 Ubiquitination
K20 Ubiquitination
K30 Ubiquitination
S34 Phosphorylation
Y38 Phosphorylation
K39 Ubiquitination
T81 Phosphorylation
K111 Ubiquitination
Y122 Phosphorylation
C124 S-Nitrosylation
S126 Phosphorylation
S128 Phosphorylation
S131 Phosphorylation
S141 Phosphorylation
K142 Ubiquitination
K146 Ubiquitination
K156 Ubiquitination
K160 Ubiquitination
Y177 Phosphorylation
Y178 Phosphorylation
K185 Ubiquitination
K191 Ubiquitination
S242 Phosphorylation
K277 Ubiquitination
K285 Ubiquitination
K290 Ubiquitination
K298 Ubiquitination
S302 Phosphorylation
T306 Phosphorylation
T309 Phosphorylation Q15118 (PDK1) , O15530 (PDPK1) , P78527 (PRKDC)
T313 Phosphorylation
Y316 Phosphorylation
Y327 Phosphorylation
K378 Acetylation
K378 Ubiquitination
S379 Phosphorylation
K401 Ubiquitination
K427 Ubiquitination
Y438 Phosphorylation
S447 Phosphorylation
T449 Phosphorylation
T451 Phosphorylation
Y456 Phosphorylation
S458 Phosphorylation
S474 Phosphorylation O15530 (PDPK1) , P78527 (PRKDC)
Y475 Phosphorylation
S476 Phosphorylation
S478 Phosphorylation
Site PTM Type Enzyme
S2 Acetylation
K14 Ubiquitination
S34 Phosphorylation
S120 Phosphorylation
S123 Phosphorylation
K156 Ubiquitination
K181 Ubiquitination
S197 Phosphorylation
K273 Sumoylation
K273 Ubiquitination
K281 Ubiquitination
K286 Ubiquitination
T298 Phosphorylation
T302 Phosphorylation
T305 Phosphorylation Q15118 (PDK1) , O15530 (PDPK1) , Q05513 (PRKCZ)
T309 Phosphorylation
Y312 Phosphorylation
Y323 Phosphorylation
T432 Phosphorylation
Y434 Phosphorylation
T440 Phosphorylation
T447 Phosphorylation
S472 Phosphorylation Q05513 (PRKCZ)
Y473 Phosphorylation
S474 Phosphorylation
S476 Phosphorylation
P31749/P31751/Q9Y243 as PTM Enzyme
Substrate Site Source
O14492 (SH2B2) S598 Uniprot
O14492-2 (SH2B2) S641 Uniprot
O14745 (SLC9A3R1) T156 Uniprot
O14746 (TERT) S227 Uniprot
O14746 (TERT) S824 Uniprot
O14757-1 (CHEK1) S280 Uniprot
O15111 (CHUK) T23 Uniprot
O15151-1 (MDM4) S342 Uniprot
O15151 (MDM4) S367 Uniprot
O15360-1 (FANCA) S1149 Uniprot
O15519 (CFLAR) S273 Uniprot
O43464 (HTRA2) S212 Uniprot
O43521 (BCL2L11) S87 Uniprot
O43524-1 (FOXO3) T32 Uniprot
O43524 (FOXO3) S253 Uniprot
O43524 (FOXO3) S315 Uniprot
O60285 (NUAK1) S600 Uniprot
O60331 (PIP5K1C) S555 Uniprot
O60343 (TBC1D4) S318 Uniprot
O60343 (TBC1D4) S588 Uniprot
O60343 (TBC1D4) T642 Uniprot
O60825 (PFKFB2) S466 Uniprot
O60825 (PFKFB2) S483 Uniprot
O75376 (NCOR1) S1450 Uniprot
O95999 (BCL10) S218 Uniprot
O95999 (BCL10) S231 Uniprot
P00533 (EGFR) S229 Uniprot
P02545 (LMNA) S301 Uniprot
P02545 (LMNA) S404 Uniprot
P03372 (ESR1) S104 Uniprot
P03372 (ESR1) S106 Uniprot
P03372 (ESR1) S118 Uniprot
P03372-1 (ESR1) S167 Uniprot
P03372 (ESR1) S305 Uniprot
P04049-1 (RAF1) S259 Uniprot
P04150 (NR3C1) S134 Uniprot
P04792 (HSPB1) S82 Uniprot
P05106 (ITGB3) T767 Uniprot
P05106 (ITGB3) T779 Uniprot
P05455 (SSB) T302 Uniprot
P07550 (ADRB2) S345 Uniprot
P07550 (ADRB2) S346 Uniprot
P08047 (SP1) S42 Uniprot
P08047 (SP1) T679 Uniprot
P08047 (SP1) S698 Uniprot
P08670 (VIM) S39 Uniprot
P08727 (KRT19) S35 Uniprot
P09601 (HMOX1) S188 Uniprot
P09651 (HNRNPA1) S199 Uniprot
P10275-1 (AR) S208 Uniprot
P10275-1 (AR) S215 Uniprot
P10275-2 (AR) S260 Uniprot
P10275 (AR) S792 Uniprot
P10276 (RARA) S96 Uniprot
P12755 (SKI) T458 Uniprot
P14598 (NCF1) S304 Uniprot
P14598 (NCF1) S328 Uniprot
P15056 (BRAF) S365 Uniprot
P15056 (BRAF) S429 Uniprot
P15976 (GATA1) S310 Uniprot
P16220 (CREB1) S133 Uniprot
P17302 (GJA1) S369 Uniprot
P17302 (GJA1) S373 Uniprot
P17542 (TAL1) T90 Uniprot
P18031 (PTPN1) S50 Uniprot
P19174 (PLCG1) S1248 Uniprot
P19634 (SLC9A1) S648 Uniprot
P19634 (SLC9A1) S703 Uniprot
P19634 (SLC9A1) S796 Uniprot
P21333 (FLNA) S2152 Uniprot
P21453 (S1PR1) T236 Uniprot
P22736 (NR4A1) S351 Uniprot
P23396 (RPS3) T70 Uniprot
P23588 (EIF4B) S422 Uniprot
P23769-1 (GATA2) S401 Uniprot
P24941 (CDK2) T39 Uniprot
P26358 (DNMT1) S143 Uniprot
P26678 (PLN) S16 Uniprot
P29317 (EPHA2) S897 Uniprot
P29401 (TKT) T382 Uniprot
P29401 (TKT) S387 Uniprot
P29474 (NOS3) S615 Uniprot
P29474-1 (NOS3) S1177 Uniprot
P29474 (NOS3) S1179 Uniprot
P30291 (WEE1) S642 Uniprot
P30305 (CDC25B) S353 Uniprot
P31749 (AKT1) T72 Uniprot
P31749 (AKT1) S246 Uniprot
P31749 (AKT1) T308 Uniprot
P31749 (AKT1) S473 Uniprot
P33778 (HIST1H2BB) S37 Uniprot
P35222 (CTNNB1) S552 Uniprot
P35226 (BMI1) S316 Uniprot
P35232 (PHB) T258 Uniprot
P35240 (NF2) S10 Uniprot
P35240 (NF2) T230 Uniprot
P35240 (NF2) S315 Uniprot
P35568 (IRS1) S629 Uniprot
P38398-8 (BRCA1) T462 Uniprot
P38398 (BRCA1) T509 Uniprot
P38398 (BRCA1) S694 Uniprot
P38936 (CDKN1A) T145 Uniprot
P38936 (CDKN1A) S146 Uniprot
P41219 (PRPH) S59 Uniprot
P41279 (MAP3K8) S400 Uniprot
P41279-1 (MAP3K8) S413 Uniprot
P42345 (MTOR) T2446 Uniprot
P42345 (MTOR) S2448 Uniprot
P42858 (HTT) S419 Uniprot
P45985-1 (MAP2K4) S80 Uniprot
P46527 (CDKN1B) S10 Uniprot
P46527 (CDKN1B) T157 Uniprot
P46527 (CDKN1B) T187 Uniprot
P46527 (CDKN1B) T198 Uniprot
P46937 (YAP1) S127 Uniprot
P47870 (GABRB2) S472 Uniprot
P48431 (SOX2) T118 Uniprot
P48730 (CSNK1D) S370 Uniprot
P49760 (CLK2) S34 Uniprot
P49760 (CLK2) T127 Uniprot
P49815 (TSC2) S939 Uniprot
P49815 (TSC2) S981 Uniprot
P49815-1 (TSC2) T1462 Uniprot
P49840 (GSK3A) S21 Uniprot
P49841 (GSK3B) S9 Uniprot
P49918 (CDKN1C) S282 Uniprot
P49918 (CDKN1C) T310 Uniprot
P51617-4 (IRAK1) T66 Uniprot
P51617 (IRAK1) T100 Uniprot
P53365 (ARFIP2) S260 Uniprot
P53804 (TTC3) S378 Uniprot
P54253 (ATXN1) S775 Uniprot
P54274 (TERF1) T273 Uniprot
P54578 (USP14) S432 Uniprot
P54646 (PRKAA2) S491 Uniprot
P55072 (VCP) S352 Uniprot
P55072 (VCP) S746 Uniprot
P55072 (VCP) S748 Uniprot
P55211-1 (CASP9) S183 Uniprot
P55211 (CASP9) S196 Uniprot
P55316 (FOXG1) T279 Uniprot
P56270 (MAZ) T385 Uniprot
P60484-1 (PTEN) S380 Uniprot
P60484-1 (PTEN) S385 Uniprot
P62136 (PPP1CA) T320 Uniprot
P63000 (RAC1) S71 Uniprot
P63104-1 (YWHAZ) S58 Uniprot
P63104-1 (YWHAZ) S184 Uniprot
P67809 (YBX1) S102 Uniprot
P68400 (CSNK2A1) T13 Uniprot
P78362 (SRPK2) T492 Uniprot
P78371 (CCT2) S260 Uniprot
P84022 (SMAD3) T179 Uniprot
P84243 (H3F3B) S11 Uniprot
P98170 (XIAP) S87 Uniprot
P98177 (FOXO4) T32 Uniprot
P98177 (FOXO4) S197 Uniprot
P98177 (FOXO4) S262 Uniprot
P98177-1 (FOXO4) T451 Uniprot
P98177-1 (FOXO4) T455 Uniprot
Q00987 (MDM2) S166 Uniprot
Q00987-11 (MDM2) S172 Uniprot
Q00987 (MDM2) S186 Uniprot
Q00987 (MDM2) S188 Uniprot
Q00987-11 (MDM2) S192 Uniprot
Q01860 (POU5F1) T235 Uniprot
Q04912 (MST1R) S1394 Uniprot
Q05195 (MXD1) S145 Uniprot
Q05397 (PTK2) S695 Uniprot
Q05397 (PTK2) T700 Uniprot
Q06187 (BTK) S51 Uniprot
Q06187 (BTK) T495 Uniprot
Q07352 (ZFP36L1) S92 Uniprot
Q07352 (ZFP36L1) S203 Uniprot
Q07812 (BAX) S184 Uniprot
Q09472 (EP300) S1834 Uniprot
Q12778 (FOXO1) T24 Uniprot
Q12778 (FOXO1) S256 Uniprot
Q12778 (FOXO1) S319 Uniprot
Q12906 (ILF3) S647 Uniprot
Q13009 (TIAM1) S231 Uniprot
Q13043 (STK4) T120 Uniprot
Q13043 (STK4) T387 Uniprot
Q13107 (USP4) S445 Uniprot
Q13131 (PRKAA1) S496 Uniprot
Q13153 (PAK1) S21 Uniprot
Q13188 (STK3) T117 Uniprot
Q13188 (STK3) T384 Uniprot
Q13309 (SKP2) S72 Uniprot
Q13322 (GRB10) S428 Uniprot
Q13370-1 (PDE3B) S295 Uniprot
Q13370-1 (PDE3B) S318 Uniprot
Q13370-1 (PDE3B) S442 Uniprot
Q13541 (EIF4EBP1) T37 Uniprot
Q13541 (EIF4EBP1) T46 Uniprot
Q13541 (EIF4EBP1) S65 Uniprot
Q13950 (RUNX2) S28 Uniprot
Q13950 (RUNX2) S196 Uniprot
Q13950 (RUNX2) T198 Uniprot
Q13950 (RUNX2) T200 Uniprot
Q14315 (FLNC) S2233 Uniprot
Q14457 (BECN1) S295 Uniprot
Q14643 (ITPR1) S2690 Uniprot
Q14957 (GRIN2C) S1081 Uniprot
Q15027 (ACAP1) S554 Uniprot
Q15121 (PEA15) S116 Uniprot
Q15365 (PCBP1) S43 Uniprot
Q15672 (TWIST1) S42 Uniprot
Q15672 (TWIST1) S123 Uniprot
Q15910-2 (EZH2) S21 Uniprot
Q15942 (ZYX) S142 Uniprot
Q16584 (MAP3K11) S674 Uniprot
Q16763 (UBE2S) T152 Uniprot
Q16875 (PFKFB3) S461 Uniprot
Q3V6T2-2 (CCDC88A) S1417 Uniprot
Q53EL6 (PDCD4) S67 Uniprot
Q53EL6 (PDCD4) S457 Uniprot
Q5VWQ8 (DAB2IP) S971 Uniprot
Q6R327 (RICTOR) T1135 Uniprot
Q7L5N1 (COPS6) S60 Uniprot
Q7Z589 (EMSY) S209 Uniprot
Q7Z5H3 (ARHGAP22) S16 Uniprot
Q7Z6J0 (SH3RF1) S304 Uniprot
Q86TI0 (TBC1D1) S235 Uniprot
Q86TI0 (TBC1D1) T596 Uniprot
Q86V81 (ALYREF) S34 Uniprot
Q86V81 (ALYREF) T219 Uniprot
Q86VP3 (PACS2) S437 Uniprot
Q86YN6 (PPARGC1B) S570 Uniprot
Q8IYJ3 (SYTL1) S241 Uniprot
Q8N6T7 (SIRT6) S338 Uniprot
Q8NCD3 (HJURP) S486 Uniprot
Q8WX93 (PALLD) S1118 Uniprot
Q8WYL5 (SSH1) T826 Uniprot
Q92547 (TOPBP1) S1159 Uniprot
Q92793 (CREBBP) T1871 Uniprot
Q92794 (KAT6A) T369 Uniprot
Q92879 (CELF1) S28 Uniprot
Q92934 (BAD) S75 Uniprot
Q92934 (BAD) S99 Uniprot
Q92934 (BAD) S118 Uniprot
Q92945 (KHSRP) S193 Uniprot
Q96B36 (AKT1S1) T246 Uniprot
Q96F86 (EDC3) S161 Uniprot
Q96HP0 (DOCK6) S1194 Uniprot
Q96QB1-1 (DLC1) S327 Uniprot
Q96QB1-1 (DLC1) S329 Uniprot
Q96QB1-1 (DLC1) S567 Uniprot
Q96QB1-2 (DLC1) S764 Uniprot
Q96QB1-2 (DLC1) S766 Uniprot
Q96QB1-2 (DLC1) S1004 Uniprot
Q96S44 (TP53RK) S250 Uniprot
Q99418 (CYTH2) T276 Uniprot
Q99490-2 (AGAP2) S472 Uniprot
Q99490-2 (AGAP2) S629 Uniprot
Q99490 (AGAP2) S985 Uniprot
Q99502 (EYA1) S299 Uniprot
Q99623 (PHB2) S91 Uniprot
Q99640 (PKMYT1) S83 Uniprot
Q99683 (MAP3K5) S83 Uniprot
Q99697 (PITX2) T90 Uniprot
Q9BPZ7 (MAPKAP1) T86 Uniprot
Q9BRQ0 (PYGO2) S48 Uniprot
Q9BVI0 (PHF20) S265 Uniprot
Q9BVI0 (PHF20) S291 Uniprot
Q9BWT1 (CDCA7) T163 Uniprot
Q9BZQ8 (FAM129A) S602 Uniprot
Q9GZV1 (ANKRD2) S99 Uniprot
Q9H3Z4 (DNAJC5) S10 Uniprot
Q9H4A3 (WNK1) T60 Uniprot
Q9H4X1-2 (RGCC) S45 Uniprot
Q9H6Z4 (RANBP3) S126 Uniprot
Q9H9Q4 (NHEJ1) T181 Uniprot
Q9HBH7 (BEX1) S102 Uniprot
Q9NR28 (DIABLO) S67 Uniprot
Q9NWV8 (BABAM1) S29 Uniprot
Q9NZJ5 (EIF2AK3) T802 Uniprot
Q9P0V3 (SH3BP4) S246 Uniprot
Q9UBK2 (PPARGC1A) S571 Uniprot
Q9UBP6 (METTL1) S27 Uniprot
Q9UKV3 (ACIN1) S1180 Uniprot
Q9UKV3 (ACIN1) S1329 Uniprot
Q9UN36 (NDRG2) S332 Uniprot
Q9UN36 (NDRG2) T348 Uniprot
Q9UQB3 (CTNND2) T457 Uniprot
Q9UQC2 (GAB2) S159 Uniprot
Q9Y261 (FOXA2) T156 Uniprot
Q9Y2I7 (PIKFYVE) S307 Uniprot
Q9Y2I7 (PIKFYVE) S318 Uniprot
Q9Y2V2 (CARHSP1) S52 Uniprot
Q9Y3C5 (RNF11) T135 Uniprot
Q9Y3M2 (CBY1) S20 Uniprot
Substrate Site Source
O15111 (CHUK) T23 Uniprot
O43464 (HTRA2) S212 Uniprot
O43524 (FOXO3) T32 Uniprot
O43524 (FOXO3) S253 Uniprot
O43524 (FOXO3) S315 Uniprot
O60331 (PIP5K1C) S555 Uniprot
P03372-1 (ESR1) S167 Uniprot
P04406 (GAPDH) T237 Uniprot
P04792 (HSPB1) S82 Uniprot
P15056 (BRAF) S364 Uniprot
P15056 (BRAF) S428 Uniprot
P15056 (BRAF) T440 Uniprot
P15311 (EZR) T567 Uniprot
P16220 (CREB1) S133 Uniprot
P28906 (CD34) S346 Uniprot
P29474 (NOS3) S615 Uniprot
P30405 (PPIF) S31 Uniprot
P35222 (CTNNB1) S552 Uniprot
P46527 (CDKN1B) T157 Uniprot
P49760 (CLK2) T344 Uniprot
P49815 (TSC2) S939 Uniprot
P49815 (TSC2) T1462 Uniprot
P49841 (GSK3B) S9 Uniprot
P55211 (CASP9) S196 Uniprot
P68431 (HIST1H3J) S11 Uniprot
P68431 (HIST1H3J) S29 Uniprot
P68431 (HIST1H3J) T46 Uniprot
P98170 (XIAP) S87 Uniprot
P98177 (FOXO4) T32 Uniprot
Q00987 (MDM2) S166 Uniprot
Q00987 (MDM2) S186 Uniprot
Q01860 (POU5F1) T235 Uniprot
Q09472 (EP300) S1834 Uniprot
Q12778 (FOXO1) T24 Uniprot
Q12778 (FOXO1) S256 Uniprot
Q13043 (STK4) T387 Uniprot
Q13188 (STK3) T117 Uniprot
Q13243 (SRSF5) S86 Uniprot
Q13541 (EIF4EBP1) T36 Uniprot
Q13541 (EIF4EBP1) T45 Uniprot
Q14315 (FLNC) S2233 Uniprot
Q15365 (PCBP1) S43 Uniprot
Q6ZWJ1 (STXBP4) S99 Uniprot
Q7Z6J0 (SH3RF1) S304 Uniprot
Q86YS7 (C2CD5) S197 Uniprot
Q8WX93 (PALLD) S1118 Uniprot
Q92908-2 (GATA6) S290 Uniprot
Q92934 (BAD) S99 Uniprot
Q92934 (BAD) S118 Uniprot
Q92945 (KHSRP) S193 Uniprot
Q96B36 (AKT1S1) T246 Uniprot
Q96F86 (EDC3) S161 Uniprot
Q99683 (MAP3K5) S83 Uniprot
Q99697 (PITX2) T90 Uniprot
Q9GZV1 (ANKRD2) S99 Uniprot
Q9H0H5 (RACGAP1) T249 Uniprot
Q9H0K1 (SIK2) S358 Uniprot
Q9NZJ5 (EIF2AK3) T802 Uniprot
Q9UBK2 (PPARGC1A) S571 Uniprot
Q9Y3M2 (CBY1) S20 Uniprot
Q9Y4I1 (MYO5A) T1650 Uniprot
Substrate Site Source
O15111 (CHUK) T23 Uniprot
O15119 (TBX3) S719 Uniprot
O43524 (FOXO3) T32 Uniprot
O43524 (FOXO3) S253 Uniprot
O43524 (FOXO3) S315 Uniprot
P04792 (HSPB1) S82 Uniprot
P15056 (BRAF) S364 Uniprot
P15056 (BRAF) S365 Uniprot
P15056 (BRAF) S428 Uniprot
P15056 (BRAF) S429 Uniprot
P16220 (CREB1) S133 Uniprot
P29474 (NOS3) S615 Uniprot
P49841 (GSK3B) S9 Uniprot
P55211 (CASP9) S196 Uniprot
Q01860 (POU5F1) T235 Uniprot
Q13043 (STK4) T387 Uniprot
Q13188 (STK3) T117 Uniprot
Q92934 (BAD) S99 Uniprot
Q92934 (BAD) S118 Uniprot
Q9UKV8 (AGO2) S387 Uniprot
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.