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RB Transcriptional Corepressor 1 Peptide 3

Antibody: CPTC-RB1-5
Background

Catalog Number:

CPTC-RB1-5

Target Antigen:

RB Transcriptional Corepressor 1 Peptide 3

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

11/29/2022

Antigen Recognition(s):

Peptide

Characterization Data [Compare Characterization Data]
  Affinity Measurement by Biolayer Interferometry
Click to enlarge image The affinity and binding kinetics of CPTC-RB1-5 antibody and BSA-conjugated phosphorylated peptide “TAVIPING-pS-PR" were measured using biolayer interferometry. BSA-conjugated peptide was covalently immobilized onto AR2G biosensors using standard amine coupling. CPTC-RB1-5 antibody, 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1 nM and 0.25 nM, was used as analyte. Buffer only and biosensors immobilized without BSA-conjugated peptide were used as references for background subtraction. All data was analyzed globally using a 1:2 bivalent model. Click image to enlarge

CPTC-RB1-5 Affinity and Kinetics (Bio-Layer Interferometry)

Result: Positive

The affinity and binding kinetics of CPTC-RB1-5 antibody and BSA-conjugated phosphorylated peptide “TAVIPING-pS-PR" were measured using biolayer interferometry. BSA-conjugated peptide was covalently immobilized onto AR2G biosensors using standard amine coupling. CPTC-RB1-5 antibody, 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1 nM and 0.25 nM, was used as analyte. Buffer only and biosensors immobilized without BSA-conjugated peptide were used as references for background subtraction. All data was analyzed globally using a 1:2 bivalent model.


Characterization SOP Files

  Affinity Measurement by SPR
Click to enlarge image The affinity and binding kinetics of CPTC-RB1-5 antibody and BSA-conjugated phosphorylated-peptide, “TAVIPING-pS-PR” (A) and non-phosphorylated peptide (B), were measured using surface plasmon resonance. Peptide was amine coupled onto a Series S CM5 biosensor chip. Antibody at 64 nM, 16 nM, 4 nM, 1.0 nM, 0.25 nM and 0.0625 nM, was used as analyte. Binding data were double-referenced and analyzed globally using a bivalent fitting model Click image to enlarge

CPTC-RB1-5 Affinity and Kinetics (Surface Plasmon Resonance) 

Result: Positive

The affinity and binding kinetics of CPTC-RB1-5 antibody and BSA-conjugated phosphorylated-peptide, “TAVIPING-pS-PR” (A) and non-phosphorylated peptide (B), were measured using surface plasmon resonance. Peptide was amine coupled onto a Series S CM5 biosensor chip. Antibody at 64 nM, 16 nM, 4 nM, 1.0 nM, 0.25 nM and 0.0625 nM, was used as analyte. Binding data were double-referenced and analyzed globally using a bivalent fitting model


Characterization SOP Files

  Immuno-MRM
Click to enlarge image Immuno-MRM using CPTC-RB1-5 as capture antibody against the phosphorylated synthetic peptide TAVIPING(pS)PR (phosphosite S249) and the correspondent non-phosphorylated peptide TAVIPINGSPR. Antibody CPTC-RB1-5 captures specifically the phosphorylated synthetic peptide, but not the correspondent non-phosphorylated peptide. Click image to enlarge

CPTC-RB1-5 iMRM (synthetic peptide)

Result: Positive

Immuno-MRM using CPTC-RB1-5 as capture antibody against the phosphorylated synthetic peptide TAVIPING(pS)PR (phosphosite S249) and the correspondent non-phosphorylated peptide TAVIPINGSPR. Antibody CPTC-RB1-5 captures specifically the phosphorylated synthetic peptide, but not the correspondent non-phosphorylated peptide.


Characterization SOP Files

  Indirect ELISA
Click to enlarge image Indirect ELISA using CPTC-RB1-5 as primary antibody against BSA conjugated phosphorylated peptide, "TAVIPING-pS-PR", coated on the plate and detected using HRP conjugated goat anti-rabbit secondary antibody and TMB. Click image to enlarge

CPTC-RB1-5 Indirect ELISA

Result: Positive

Indirect ELISA using CPTC-RB1-5 as primary antibody against BSA conjugated phosphorylated peptide, "TAVIPING-pS-PR", coated on the plate and detected using HRP conjugated goat anti-rabbit secondary antibody and TMB.


Characterization SOP Files

  NCI 60 Protein Array

CPTC-RB1-5 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western blot using CPTC-RB1-5 as primary antibody against whole cell lysates CCRF-CEM (lane 2), HeLa (lane 3), Jurkat (lane 4), K-562 (lane 5), and MCF7 (lane 6). The expected molecular weight is 106.2 kDa. All cell lines are negative. Click image to enlarge

CPTC-RB1-5 Western Blot (Cell Lysates)

Result: Negative

Western blot using CPTC-RB1-5 as primary antibody against whole cell lysates CCRF-CEM (lane 2), HeLa (lane 3), Jurkat (lane 4), K-562 (lane 5), and MCF7 (lane 6). The expected molecular weight is 106.2 kDa. All cell lines are negative.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated western blot using CPTC-RB1-5 as primary antibody against whole cell lysates of cell lines CCRF-CEM, HeLa, Jurkat, K-562 and MCF7. Protein molecular weight is about 106 KDa. The antibody cannot recognize the target in any of the tested lysates. Loading controls were run with anti-GAPDH antibody. Click image to enlarge

CPTC-RB1-5 Automated Western Blot (Cell Lysates)

Result: Negative

Automated western blot using CPTC-RB1-5 as primary antibody against whole cell lysates of cell lines CCRF-CEM, HeLa, Jurkat, K-562 and MCF7. Protein molecular weight is about 106 KDa. The antibody cannot recognize the target in any of the tested lysates. Loading controls were run with anti-GAPDH antibody.


Characterization SOP Files

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Antigen: RB Transcriptional Corepressor 1 Peptide 3
Background

NCI Identification Number:

00528

Antigen Name:

RB Transcriptional Corepressor 1 Peptide 3

CPTC Name:

CPTC-RB1 Peptide 3

Aliases:

RB Transcriptional Corepressor 1; PPP1R130; RB; Protein Phosphatase 1, Regulatory Subunit 130; Prepro-Retinoblastoma-Associated Protein; Retinoblastoma-Associated Protein; Retinoblastoma 1; P110-RB1; P105-Rb; Pp110; OSRC; PRb; Exon 17 Tumor GOS561 Substitution Mutation Causes Premature Stop; GOS563 Exon 17 Substitution Mutation Causes Premature Stop; Retinoblastoma Suspectibility Protein; Osteosarcoma; Rb

Function:

The protein encoded by this gene is a negative regulator of the cell cycle and was the first tumor suppressor gene found. The encoded protein also stabilizes constitutive heterochromatin to maintain the overall chromatin structure. The active, hypophosphorylated form of the protein binds transcription factor E2F1. Defects in this gene are a cause of childhood cancer retinoblastoma (RB), bladder cancer, and osteogenic sarcoma. [provided by RefSeq, Jul 2008]RB1 (RB Transcriptional Corepressor 1) is a Protein Coding gene. Diseases associated with RB1 include Retinoblastoma and Small Cell Cancer Of The Lung. Among its related pathways are 4-1BB Pathway and Aberrant regulation of mitotic G1/S transition in cancer due to RB1 defects. Gene Ontology (GO) annotations related to this gene include DNA-binding transcription factor activity and enzyme binding. An important paralog of this gene is RBL2.Tumor suppressor that is a key regulator of the G1/S transition of the cell cycle (PubMed:10499802). The hypophosphorylated form binds transcription regulators of the E2F family, preventing transcription of E2F-responsive genes (PubMed:10499802). Both physically blocks E2Fs transactivating domain and recruits chromatin-modifying enzymes that actively repress transcription (PubMed:10499802). Cyclin and CDK-dependent phosphorylation of RB1 induces its dissociation from E2Fs, thereby activating transcription of E2F responsive genes and triggering entry into S phase (PubMed:10499802). RB1 also promotes the G0-G1 transition upon phosphorylation and activation by CDK3/cyclin-C (PubMed:15084261). 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). RB_HUMAN,P06400(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. RB_HUMAN,P06400

Chromosomal Localization:

13q14.2

Accession Number:

NP_000312.2

UniProt Accession Number:

P06400

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

TAVIPING-pS-PR

Native Sequence:

Calculated Isoelectric Point:

Molecular Weight:

1210

Last Updated:

06/09/2022

Links

Characterization Data

SOPs

No SOPs available.

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