Phospho-Retinoblastoma (Ser780) Antibody

Product:	Phospho-Retinoblastoma (Ser780) Antibody
Catalog:	AF3103
Description:	Rabbit polyclonal antibody to Phospho-Retinoblastoma (Ser780)
Application:	WB IHC IF/ICC
Cited expt.:	WB, IHC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.:	106kDa; 106kD(Calculated).
Uniprot:	P06400
RRID:	AB_2834540

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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. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
100ul	$280	In stock
200ul	$350	In stock

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Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

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:

Pig(100%), Bovine(100%), Horse(91%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(91%), Xenopus(90%)

Clonality:

Polyclonal

Specificity:

Phospho-Retinoblastoma (Ser780) Antibody detects endogenous levels of Retinoblastoma only when phosphorylated at Serine 780.

RRID:

AB_2834540
Cite Format: Affinity Biosciences Cat# AF3103, RRID:AB_2834540.

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

Exon 17 tumor GOS561 substitution mutation causes premature stop; GOS563 exon 17 substitution mutation causes premature stop; OSRC; Osteosarcoma; p105-Rb; P105RB; PP105; pp110; PPP1R130; pRb; Prepro retinoblastoma associated protein; Protein phosphatase 1 regulatory subunit 130; Rb; RB transcriptional corepressor 1; RB_HUMAN; RB1; RB1 gene; Retinoblastoma 1; Retinoblastoma suspectibility protein; Retinoblastoma-associated protein;

Immunogens

Immunogen:

A synthesized peptide derived from human Retinoblastoma around the phosphorylation site of Ser780.

Uniprot:

P06400(RB_HUMAN) >>Visit HPA database.

Gene(ID):

RB1(5925)

Expression:

P06400 RB_HUMAN:

Expressed in the retina. Expressed in foreskin keratinocytes (at protein level) (PubMed:20940255).

Description:

Retinoblastoma (RB) is an embryonic malignant neoplasm of retinal origin. It almost always presents in early childhood and is often bilateral. Spontaneous regression ('cure') occurs in some cases.

Sequence:

P06400 RB_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MPPKTPRKTAATAAAAAAEPPAPPPPPPPEEDPEQDSGPEDLPLVRLEFEETEEPDFTALCQKLKIPDHVRERAWLTWEKVSSVDGVLGGYIQKKKELWGICIFIAAVDLDEMSFTFTELQKNIEISVHKFFNLLKEIDTSTKVDNAMSRLLKKYDVLFALFSKLERTCELIYLTQPSSSISTEINSALVLKVSWITFLLAKGEVLQMEDDLVISFQLMLCVLDYFIKLSPPMLLKEPYKTAVIPINGSPRTPRRGQNRSARIAKQLENDTRIIEVLCKEHECNIDEVKNVYFKNFIPFMNSLGLVTSNGLPEVENLSKRYEEIYLKNKDLDARLFLDHDKTLQTDSIDSFETQRTPRKSNLDEEVNVIPPHTPVRTVMNTIQQLMMILNSASDQPSENLISYFNNCTVNPKESILKRVKDIGYIFKEKFAKAVGQGCVEIGSQRYKLGVRLYYRVMESMLKSEEERLSIQNFSKLLNDNIFHMSLLACALEVVMATYSRSTSQNLDSGTDLSFPWILNVLNLKAFDFYKVIESFIKAEGNLTREMIKHLERCEHRIMESLAWLSDSPLFDLIKQSKDREGPTDHLESACPLNLPLQNNHTAADMYLSPVRSPKKKGSTTRVNSTANAETQATSAFQTQKPLKSTSLSLFYKKVYRLAYLRLNTLCERLLSEHPELEHIIWTLFQHTLQNEYELMRDRHLDQIMMCSMYGICKVKNIDLKFKIIVTAYKDLPHAVQETFKRVLIKEEEYDSIIVFYNSVFMQRLKTNILQYASTRPPTLSPIPHIPRSPYKFPSSPLRIPGGNIYISPLKSPYKISEGLPTPTKMTPRSRILVSIGESFGTSEKFQKINQMVCNSDRVLKRSAEGSNPPKPLKKLRFDIEGSDEADGSKHLPGESKFQQKLAEMTSTRTRMQKQKMNDSMDTSNKEEK

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

Pig

100

Bovine

100

Sheep

100

Dog

100

Rabbit

100

Horse

Chicken

Xenopus

Zebrafish

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

P06400_RB_HUMAN

Function:

Key regulator of entry into cell division that acts as a tumor suppressor. Promotes G0-G1 transition when phosphorylated by CDK3/cyclin-C. Acts as a transcription repressor of E2F1 target genes. The underphosphorylated, active form of RB1 interacts with E2F1 and represses its transcription activity, leading to cell cycle arrest. Directly involved in heterochromatin formation by maintaining overall chromatin structure and, in particular, that of constitutive heterochromatin by stabilizing histone methylation. Recruits and targets histone methyltransferases SUV39H1, KMT5B and KMT5C, leading to epigenetic transcriptional repression. Controls histone H4 'Lys-20' trimethylation. Inhibits the intrinsic kinase activity of TAF1. Mediates transcriptional repression by SMARCA4/BRG1 by recruiting a histone deacetylase (HDAC) complex to the c-FOS promoter. In resting neurons, transcription of the c-FOS promoter is inhibited by BRG1-dependent recruitment of a phospho-RB1-HDAC1 repressor complex. Upon calcium influx, RB1 is dephosphorylated by calcineurin, which leads to release of the repressor complex (By similarity).

(Microbial infection) In case of viral infections, interactions with SV40 large T antigen, HPV E7 protein or adenovirus E1A protein induce the disassembly of RB1-E2F1 complex thereby disrupting RB1's activity.

PTMs:

Phosphorylated by CDK6 and CDK4, and subsequently by CDK2 at Ser-567 in G1, thereby releasing E2F1 which is then able to activate cell growth. Dephosphorylated at the late M phase. SV40 large T antigen, HPV E7 and adenovirus E1A bind to the underphosphorylated, active form of pRb. Phosphorylation at Thr-821 and Thr-826 promotes interaction between the C-terminal domain C and the Pocket domain, and thereby inhibits interactions with heterodimeric E2F/DP transcription factor complexes. Dephosphorylated at Ser-795 by calcineruin upon calcium stimulation. CDK3/cyclin-C-mediated phosphorylation at Ser-807 and Ser-811 is required for G0-G1 transition. Phosphorylated by CDK1 and CDK2 upon TGFB1-mediated apoptosis (By similarity).

N-terminus is methylated by METTL11A/NTM1 (By similarity). Monomethylation at Lys-810 by SMYD2 enhances phosphorylation at Ser-807 and Ser-811, and promotes cell cycle progression. Monomethylation at Lys-860 by SMYD2 promotes interaction with L3MBTL1.

Acetylated during keratinocyte differentiation. Acetylation at Lys-873 and Lys-874 regulates subcellular localization. Can be deacetylated by SIRT1.

Subcellular Location:

Nucleus.
Note: During keratinocyte differentiation, acetylation by KAT2B/PCAF is required for nuclear localization.

Tissue Specificity:

Expressed in the retina. Expressed in foreskin keratinocytes (at protein level).

Family&Domains:

The Pocket domain binds to the threonine-phosphorylated domain C, thereby preventing interaction with heterodimeric E2F/DP transcription factor complexes.

Belongs to the retinoblastoma protein (RB) family.

Research Fields

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

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

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

· 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 > Viral carcinogenesis.

· Human Diseases > Cancers: Specific types > Pancreatic 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 > 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)

References

1). Blockage of EGFR/AKT and mevalonate pathways synergize the antitumor effect of temozolomide by reprogramming energy metabolism in glioblastoma. Cancer communications (London, England), 2023 (PubMed: 37920878) [IF=16.2]

Application: WB Species: Human Sample: TBD0220, U-87 MG, and U-87 MG-EGFR-vIII cells

FIGURE 3 EGFR/AKT pathway regulates mitochondrial respiration and proliferation in GBM cells. (A-B) TBD0220 (A), and U-87 MG-EGFR-vIII cells (B) were treated with DMSO or 5 μmol/L MK-2206 for 24 h. The mitochondrial functions were monitored by Seahorse XF Cell Mito Stress test. The OCR, basal respiration, proton leak, and ATP production rates were measured as illustrated. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (independent-sample Student's t-test for TBD0220; one-way ANOVA for U-87 MG). (C) ATP levels in TBD0220, U-87 MG, and U-87 MG-EGFR-vIII cells were analyzed after 24 h of treatments with DMSO or 5 μmol/L of MK-2206. ∗∗P < 0.01, ∗∗∗P < 0.001 (independent-sample Student's t-test for TBD0220; one-way ANOVA for U-87 MG). (D) The cell growth assay for TBD0220, U-87 MG, and U-87 MG-EGFR-vIII lines treated with DMSO, 5 μmol/L MK-2206, or 5 μmol/L MK-2206 plus 50 μmol/L ATP were performed. ∗∗∗P < 0.001 (two-way ANOVA). (E) The colony formation assay of TBD0220, U-87 MG, and U-87 MG-EGFR-vIII lines treated with DMSO or 1 μmol/L MK-2206. (F) Cell cycle distributions were analyzed by flow cytometry in TBD0220, U-87 MG, and U-87 MG-EGFR-vIII cells treated with DMSO or 5 μmol/L MK-2206. (G) Western blotting to show changes in expressions of CDK2, CDK4, CDK6, Cyclin D, RB, p-RB, and GAPDH in TBD0220, U-87 MG, and U-87 MG-EGFR-vIII cells treated with DMSO or MK-2206. Abbreviations: EGFR, epidermal growth factor receptor; AKT, AKT serine/threonine kinase 1; GBM, glioblastoma; ANOVA, analysis of variance; DMSO, dimethyl sulfoxide; CDK2, cyclin-dependent kinase 2; CDK4, cyclin-dependent kinase 4; CDK6, cyclin-dependent kinase 6; OCR, oxygen consumption rate.

2). TCA-phospholipid-glycolysis targeted triple therapy effectively suppresses ATP production and tumor growth in glioblastoma. Theranostics, 2023 (PubMed: 36276638) [IF=12.4]

Application: WB Species: Human Sample: TBD0220 and U87MG cells

Figure 3 Reduction in ATP production hinders cell proliferation and contributes to G1/S arrest in GBM. (A-B) The relative viability of TBD0220 (A) and U87MG (B) cells was measured using a CCK-8 kit (n = 3). (C-D) Colony formation assay to detect GBM cell growth (D), and the quantification of colony numbers (C) (n = 3). (E) Cell cycle analysis using flow cytometry after incubation with different treatments like EPIC (20 µM), AA (25 µM), or EPIC (20 µM) + AA (25 µM) for 48 h, and the results are plotted as a histogram (n = 3) (F) Representative western blotting showing the expression of p21 and Rb and their downstream targets. The results were normalized to Tubulin with the control group as 1. Protein expression was quantified by ImageJ. (G) Representative confocal images of CDK6 after the treatment with AA (25 µM), EPIC (20 µM), or EPIC (20 µM) + AA (25 µM) for 48 h (n = 6). (H) The quantitative analysis of fluorescence images of CDK6 (n = 6). All data are shown as the mean values ± SD, and p values are based on one-way or two-way ANOVA. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05. Scale bar = 20 μm.

3). Combination LSD1 and HOTAIR-EZH2 inhibition disrupts cell cycle processes and induces apoptosis in glioblastoma cells. PHARMACOLOGICAL RESEARCH, 2021 (PubMed: 34246782) [IF=9.1]

4). Metformin as a senostatic drug enhances the anticancer efficacy of CDK4/6 inhibitor in head and neck squamous cell carcinoma. Cell Death & Disease, 2020 (PubMed: 33116117) [IF=8.1]

Application: WB Species: Human Sample:

Fig. 2 CDK4/6 inhibitor combined with metformin promoted cell cycle arrest. A, B Flow cytometry results showed that the combination of the CDK4/6 inhibitor LY2835219 (HSC3: 0.1 μM; HSC6: 0.3 μM; Cal27: 1.25 μM) with metformin (HSC3: 10 mM; HSC6: 1 mM; Cal27: 10 mM) induced cell cycle arrest in the G0/G1 phase. C Western blot results showed that LY2835219 upregulated the expression of p16 and p21 and downregulated the expression of pRb. D Western blot results showed that metformin upregulated the expression of p16 and p21 and downregulated the expression of pRb. E Compared with LY2835219 monotherapy, the combination treatment significantly upregulated p21 expression and suppressed p-Rb expression. F Immunohistochemistry results (400×) showed that LY2835219 and metformin synergistically upregulated p21 expression and downregulated pRb expression in HSC6 xenograft tumors. Bar: 100 μm. *P

Application: IHC Species: Human Sample:

5). Combination of N, N′-dicyclohexyl-N-arachidonic acylurea and tacrolimus prolongs cardiac allograft survival in mice. Immunology and Cell Biology, 2020 (PubMed: 32162358) [IF=3.2]

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Phospho-Retinoblastoma (Ser780) Antibody - #AF3103