Product: Phospho-p21 Cip1 (Thr145) Antibody
Catalog: AF3290
Description: Rabbit polyclonal antibody to Phospho-p21 Cip1 (Thr145)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.: 21kDa,25kDa; 18kD(Calculated).
Uniprot: P38936
RRID: AB_2834711

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 100ul $280 In stock
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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Bovine(100%), Horse(91%), Sheep(100%), Rabbit(83%), Dog(100%)
Clonality:
Polyclonal
Specificity:
Phospho-p21 Cip1 (Thr145) Antibody detects endogenous levels of p21 Cip1 only when phosphorylated at Threonine 145.
RRID:
AB_2834711
Cite Format: Affinity Biosciences Cat# AF3290, RRID:AB_2834711.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
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.
Alias:

Fold/Unfold

CAP20; CDK-interacting protein 1; CDKI; CDKN1; Cdkn1a; CDN1A_HUMAN; CIP1; Cyclin Dependent Kinase Inhibitor 1A; Cyclin-dependent kinase inhibitor 1; Cyclin-dependent kinase inhibitor 1A (P21); Cyclin-dependent kinase inhibitor 1A (p21, Cip1); DNA Synthesis Inhibitor; MDA-6; MDA6; Melanoma differentiation-associated protein 6; Melanoma differentiation-associated protein; p21; P21 protein; p21CIP1; p21Cip1/Waf1; p21WAF; PIC1; SDI1; SLC12A9; WAF1; Wild type p53 activated fragment 1 (WAF1); Wild type p53 activated fragment 1; Wildtype p53-activated fragment 1;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P38936 CDN1A_HUMAN:

Expressed in all adult tissues, with 5-fold lower levels observed in the brain.

Description:
This gene encodes a potent cyclin-dependent kinase inhibitor. The encoded protein binds to and inhibits the activity of cyclin-CDK2 or -CDK4 complexes, and thus functions as a regulator of cell cycle progression at G1. The expression of this gene is tightly controlled by the tumor suppressor protein p53, through which this protein mediates the p53-dependent cell cycle G1 phase arrest in response to a variety of stress stimuli.
Sequence:
MSEPAGDVRQNPCGSKACRRLFGPVDSEQLSRDCDALMAGCIQEARERWNFDFVTETPLEGDFAWERVRGLGLPKLYLPTGPRRGRDELGGGRRPGTSPALLQGTAEEDHVDLSLSCTLVPRSGEQAEGSPGGPGDSQGRKRRQTSMTDFYHSKRRLIFSKRKP

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Bovine
100
Sheep
100
Dog
100
Horse
91
Rabbit
83
Zebrafish
67
Xenopus
56
Pig
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P38936 As Substrate

Site PTM Type Enzyme
S2 Acetylation
S15 Phosphorylation
K16 Ubiquitination
S31 Phosphorylation
T57 Phosphorylation Q16539 (MAPK14) , P28482 (MAPK1) , P45983 (MAPK8) , P49841 (GSK3B)
K75 Ubiquitination
T80 Phosphorylation
S98 Phosphorylation Q99683 (MAP3K5) , P45983 (MAPK8)
S114 Phosphorylation P49841 (GSK3B)
S116 Phosphorylation
S123 Phosphorylation
S130 Phosphorylation Q16539 (MAPK14) , P45983 (MAPK8) , P24941 (CDK2) , P28482 (MAPK1) , Q00534 (CDK6)
K141 Acetylation
K141 Ubiquitination
T145 Phosphorylation P31749 (AKT1) , Q9P1W9 (PIM2) , O14757 (CHEK1) , O43293 (DAPK3) , P11309 (PIM1)
S146 Phosphorylation P53355 (DAPK1) , O14757 (CHEK1) , P11309 (PIM1) , Q9NRM7 (LATS2) , Q9P1W9 (PIM2) , P17252 (PRKCA) , Q05655 (PRKCD) , P31749 (AKT1) , Q15208 (STK38)
Y151 Phosphorylation
S153 Phosphorylation P17252 (PRKCA) , Q9Y463 (DYRK1B)
K154 Acetylation
K154 Ubiquitination
R156 Methylation
S160 Phosphorylation
K161 Acetylation
K161 Ubiquitination
K163 Acetylation
K163 Ubiquitination

Research Backgrounds

Function:

May be involved in p53/TP53 mediated inhibition of cellular proliferation in response to DNA damage. Binds to and inhibits cyclin-dependent kinase activity, preventing phosphorylation of critical cyclin-dependent kinase substrates and blocking cell cycle progression. Functions in the nuclear localization and assembly of cyclin D-CDK4 complex and promotes its kinase activity towards RB1. At higher stoichiometric ratios, inhibits the kinase activity of the cyclin D-CDK4 complex. Inhibits DNA synthesis by DNA polymerase delta by competing with POLD3 for PCNA binding. Plays an important role in controlling cell cycle progression and DNA damage-induced G2 arrest.

PTMs:

Phosphorylation of Thr-145 by Akt or of Ser-146 by PKC impairs binding to PCNA. Phosphorylation at Ser-114 by GSK3-beta enhances ubiquitination by the DCX(DTL) complex. Phosphorylation of Thr-145 by PIM2 enhances CDKN1A stability and inhibits cell proliferation. Phosphorylation of Thr-145 by PIM1 results in the relocation of CDKN1A to the cytoplasm and enhanced CDKN1A protein stability. UV radiation-induced phosphorylation at Thr-80 by LKB1 and at Ser-146 by NUAK1 leads to its degradation.

Ubiquitinated by MKRN1; leading to polyubiquitination and 26S proteasome-dependent degradation. Ubiquitinated by the DCX(DTL) complex, also named CRL4(CDT2) complex, leading to its degradation during S phase or following UV irradiation. Ubiquitination by the DCX(DTL) complex is essential to control replication licensing and is PCNA-dependent: interacts with PCNA via its PIP-box, while the presence of the containing the 'K+4' motif in the PIP box, recruit the DCX(DTL) complex, leading to its degradation. Ubiquitination at Ser-2 leads to degradation by the proteasome pathway. Ubiquitinated by RNF114; leading to proteasomal degradation.

Acetylation leads to protein stability. Acetylated in vitro on Lys-141, Lys-154, Lys-161 and Lys-163. Deacetylation by HDAC1 is prevented by competitive binding of C10orf90/FATS to HDAC1 (By similarity).

Subcellular Location:

Cytoplasm. Nucleus.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in all adult tissues, with 5-fold lower levels observed in the brain.

Subunit Structure:

Interacts with HDAC1; the interaction is prevented by competitive binding of C10orf90/FATS to HDAC1 facilitating acetylation and protein stabilization of CDKN1A/p21 (By similarity). Interacts with MKRN1. Interacts with PSMA3. Interacts with PCNA. Component of the ternary complex, cyclin D-CDK4-CDKN1A. Interacts (via its N-terminal domain) with CDK4; the interaction promotes the assembly of the cyclin D-CDK4 complex, its nuclear translocation and promotes the cyclin D-dependent enzyme activity of CDK4. Binding to CDK2 leads to CDK2/cyclin E inactivation at the G1-S phase DNA damage checkpoint, thereby arresting cells at the G1-S transition during DNA repair. Interacts with PIM1. Interacts with STK11 and NUAK1. Interacts wih DTL. Interacts with isoform 1 and isoform 2 of TRIM39.

Family&Domains:

The PIP-box K+4 motif mediates both the interaction with PCNA and the recruitment of the DCX(DTL) complex: while the PIP-box interacts with PCNA, the presence of the K+4 submotif, recruits the DCX(DTL) complex, leading to its ubiquitination.

The C-terminal is required for nuclear localization of the cyclin D-CDK4 complex.

Belongs to the CDI family.

Research Fields

· Cellular Processes > Cell growth and death > Cell cycle.   (View pathway)

· Cellular Processes > Cell growth and death > p53 signaling pathway.   (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence.   (View pathway)

· Environmental Information Processing > Signal transduction > ErbB signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > FoxO signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Jak-STAT signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Colorectal cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Renal cell carcinoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Pancreatic cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Endometrial cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Glioma.   (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Thyroid cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Basal cell carcinoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Melanoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Bladder 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 > Cancers: Specific types > Non-small cell lung cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Breast cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer.   (View pathway)

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

References

1). Kreis et al. Functional Analysis of p21Cip1/CDKN1A and Its Family Members in Trophoblastic Cells of the Placenta and Its Roles in Preeclampsia. Cells [IF=6.0]

Application: IHC    Species: Human    Sample:

Figure 1. Cell cycle regulators are affected by the delivery mode, and expressed in trophoblast organoids and placental tissues. (A) The relative amount of the gene levels of CDKN1A (p21), CDKN1B (p27), and CDKN1C (p57) was analyzed in placental tissues from different delivery modes (n = 5). The results are presented as relative quantification (RQ) with minimum and maximum range. The mean value of the expression levels of SDHA (succinate dehydrogenase complex, subunit A), TBP (TATA box–binding protein), and YWHAZ (tyrosine 3–monooxygenase/tryptophan 5–monooxygenase activation protein, zeta polypeptide) served as the endogenous control. CS, caesarean section; eCS, emergency caesarean section after the onset of labor; VD, vaginal delivery; opVD, operative vaginal delivery. Unpaired Student’s t-test was used for statistical analysis, ** p < 0.01. (B) Representative immunohistochemistry-immunofluorescence (IHC–IF) image of a trophoblast organoid (8 weeks of gestation) stained for cytokeratin 7 (green), p21 (red), and nuclei (DAPI, blue) is shown. Scale: 50 µm. Inset scale: 10 µm. Villous cytotrophoblasts (CTBs) and the syncytiotrophoblast (STB) are indicated. (C) Representative IHC-IF image of placental tissue (8 weeks of gestation) stained for cytokeratin 7 (green), p21 (red), and nuclei (DAPI, blue) is presented. Scale: 10 µm. (D) Representative IHC–IF image of placental tissue (7 weeks of gestation) stained for E-cadherin (green), p21 (red), and nuclei (DAPI, blue) is shown. Scale: 10 µm. Inset scale: 5 µm. CTBs ongoing to fuse to the STB (fCTBs), CTBs and the STB are indicated. (E) Formalin-fixed and paraffin-embedded (FFPE) tissue sections were immunohistochemically stained with p21 (first row), p–p21 (second row), p27 (third row) or p57 (fourth row) antibody (brown), respectively, and counterstained with hematoxylin (blue). Scale: 50 µm. Inset scale: 20 µm. EVTs (extravillous cytotrophoblasts) are indicated.

2). Wang Y et al. Cholesterol regulates cell proliferation and apoptosis of colorectal cancer by modulating miR-33a-PIM3 pathway. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 2019 Apr 9;511(3):685-692 (PubMed: 30827510) [IF=3.1]

Application: WB    Species: human    Sample: CRC cells

Fig. 3.| Mechanism of miR-33a-PIM3 pathway in the role of cholesterol in CRC cells. (A) Expression of PIM3 mRNA was not changed in cholesterol treatment for 48h. (B) The changes of miR-33a after cells were transfected with miR-33a mimics or inhibitor for 24h. (C) PIM3 mRNA of cells did not show obviously difference after transfected with miR-33a mimics or inhibitor for 24h (D) PIM3 was proved to be the target gene of miR-33a by dual luciferase reporter assay. (E) The difference of expression of cell cycle and apoptotic related protein in miR-33a-PIM3 pathway between CON and CH.

3). Yang Q et al. Methyl-β-cyclodextrin potentiates the BITC-induced anti-cancer effect through modulation of the Akt phosphorylation in human colorectal cancer cells. BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 2018 Dec;82(12):2158-2167 (PubMed: 30200817) [IF=1.6]

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