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RB Transcriptional Corepressor Like 1 Peptide 1

Antibody: CPTC-RBL1-1
Background

Catalog Number:

CPTC-RBL1-1

Target Antigen:

RB Transcriptional Corepressor Like 1 Peptide 1

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

12/06/2024

Antigen Recognition(s):

Peptide, Endogenous

Characterization Data [Compare Characterization Data]
  Affinity Measurement by Biolayer Interferometry
Click to enlarge image Affinity and binding kinetics of CPTC-RBL1-1 and BSA conjugated peptide, CSIYI-pS-PHKN, were measured using biolayer interferometry. BSA-conjugated peptide was amine coupled to the AR2G biosensors. The CPTC-RBL1-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM and 0.25 nM, was used as analyte. Buffer only and biosensors immobilized without peptide were used as references for background subtraction. All binding data were analyzed globally using a 1:2 (bivalent) fitting model. Click image to enlarge

CPTC-RBL1-1 Affinity and Kinetics (Bio-Layer Interferometry) 

Result: High Binding

Affinity and binding kinetics of CPTC-RBL1-1 and BSA conjugated peptide, CSIYI-pS-PHKN, were measured using biolayer interferometry. BSA-conjugated peptide was amine coupled to the AR2G biosensors. The CPTC-RBL1-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM and 0.25 nM, was used as analyte. Buffer only and biosensors immobilized without peptide were used as references for background subtraction. All binding data were analyzed globally using a 1:2 (bivalent) fitting model.


Characterization SOP Files

  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-RBL1-1 and BSA-conjugated peptide, CSIYI-pS-PHKN, were measured using surface plasmon resonance. BSA-conjugated peptide was amine coupled onto a Series S CM5 biosensor chip. CPTC-RBL1-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1 nM, and 0.25 nM, was used as analyte. All data were double referenced and analyzed globally using a 1:2 (bivalent) fitting model. Click image to enlarge

CPTC-RBL1-1 Affinity and Kinetics (Surface Plasmon Resonance)

Result: High Binding

Affinity and binding kinetics of CPTC-RBL1-1 and BSA-conjugated peptide, CSIYI-pS-PHKN, were measured using surface plasmon resonance. BSA-conjugated peptide was amine coupled onto a Series S CM5 biosensor chip. CPTC-RBL1-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1 nM, and 0.25 nM, was used as analyte. All data were double referenced and analyzed globally using a 1:2 (bivalent) fitting model.


Characterization SOP Files

  Flow Cytometry

CPTC-RBL1-1 (Flow Cytometry)

Result: Negative

Flow cytometric analysis of thyroid medullary TT cells, asynchronous (blue) or synchronous (serum starved 24 hours, green) using CPTC-RBL1-1 rabbit antibody (solid lines) or concentration matched rabbit isotype control antibody (dashed lines). TT cells were permeabilized, fixed, and then stained with either CPTC-RBL1-1 or rabbit isotype control antibody. A BV421 conjugated goat anti-rabbit IgG (H+L) was used as a secondary antibody. All data were analyzed using FlowJo.


Characterization SOP Files

  IHC Tissue
Click to enlarge image Tissue Micro-Array (TMA) core of colon cancer showing nuclear staining using Antibody CPTC-RBL1-1. Titer: 1:6250 Click image to enlarge

CPTC-RBL1-1 IHC Tissue

Result: Positive

Tissue Micro-Array (TMA) core of colon cancer showing nuclear staining using Antibody CPTC-RBL1-1. Titer: 1:6250


Characterization SOP Files

  Indirect ELISA
Click to enlarge image Indirect ELISA using CPTC-RBL1-1 as primary antibody against BSA-conjugated peptide, CSIYI-pS-PHKN, coated on the plate and detected using the goat anti- rabbit antibody and TMB. Click image to enlarge

CPTC-RBL1-1 Indirect ELISA

Result: High Binding

Indirect ELISA using CPTC-RBL1-1 as primary antibody against BSA-conjugated peptide, CSIYI-pS-PHKN, coated on the plate and detected using the goat anti- rabbit antibody and TMB.


Characterization SOP Files

  NCI 60 Protein Array
Click to enlarge image Protein Array in which CPTC-RBL1-1 is screened against the NCI60 cell line panel for expression. Data is normalized to a mean signal of 1.0 and standard deviation of 0.5. Color conveys over-expression level (green), basal level (blue), under-expression level (red). Click image to enlarge

CPTC-RBL1-1 NCI 60 Protein Array

Result: Positive

Protein Array in which CPTC-RBL1-1 is screened against the NCI60 cell line panel for expression. Data is normalized to a mean signal of 1.0 and standard deviation of 0.5. Color conveys over-expression level (green), basal level (blue), under-expression level (red).


Characterization SOP Files

  Western Blot - Over-expressed transient protein in cell lysate

CRTP-RBL1-1 WB (Over-expressed lysates)

Result: Negative

Western Blot using CRTP-RBL1-1 as primary antibody against the over-expressed lysate of RBL1. The antibody is not able to recognize the target protein in the tested lysate.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western blot using CPTC-RBL1-1 as primary antibody against the cell lysates of TT, H146 and H1184 cells, asynchronous (+) or synchronous - serum starved 24hours (-). The antibody was able to detect (pink signal) the target protein in TT cells, and presumebly detected (presence of extra bands) in H146 and H1184 cell lysates, without any significant difference in the expression levels based on the asynchronous and synchrounous status. The same membrane was probed with an anti CytC antibody, and all the cell lysates show the presence of the housekeeping protein (green signal). Click image to enlarge

CRTP-RBL1-1 WB (Cell lysates)

Result: Positive

Western blot using CPTC-RBL1-1 as primary antibody against the cell lysates of TT, H146 and H1184 cells, asynchronous (+) or synchronous - serum starved 24hours (-). The antibody was able to detect (pink signal) the target protein in TT cells, and presumebly detected (presence of extra bands) in H146 and H1184 cell lysates, without any significant difference in the expression levels based on the asynchronous and synchrounous status. The same membrane was probed with an anti CytC antibody, and all the cell lysates show the presence of the housekeeping protein (green signal).


Characterization SOP Files

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Antigen:  RB Transcriptional Corepressor Like 1 Peptide 1
Background

NCI Identification Number:

00535

Antigen Name:

RB Transcriptional Corepressor Like 1 Peptide 1

CPTC Name:

CPTC-RBL1 Peptide 1

Aliases:

RB Transcriptional Corepressor Like 1; P107; PRB1; 107 KDa Retinoblastoma-Associated Protein; Retinoblastoma-Like Protein 1; Retinoblastoma-Like 1; Cp107; CP107; PRb1

Function:

The protein encoded by this gene is similar in sequence and possibly function to the product of the retinoblastoma 1 (RB1) gene. The RB1 gene product is a tumor suppressor protein that appears to be involved in cell cycle regulation, as it is phosphorylated in the S to M phase transition and is dephosphorylated in the G1 phase of the cell cycle. Both the RB1 protein and the product of this gene can form a complex with adenovirus E1A protein and SV40 large T-antigen, with the SV40 large T-antigen binding only to the unphosphorylated form of each protein. In addition, both proteins can inhibit the transcription of cell cycle genes containing E2F binding sites in their promoters. Due to the sequence and biochemical similarities with the RB1 protein, it is thought that the protein encoded by this gene may also be a tumor suppressor. Two transcript variants encoding different isoforms have been found for this gene.RBL1 (RB Transcriptional Corepressor Like 1) is a Protein Coding gene. Diseases associated with RBL1 include Retinoblastoma and Bilateral Retinoblastoma. Among its related pathways are Aberrant regulation of mitotic G1/S transition in cancer due to RB1 defects and G0 and Early G1. Gene Ontology (GO) annotations related to this gene include transcription factor binding. An important paralog of this gene is RBL2.Key regulator of entry into cell division (PubMed:17671431). Directly involved in heterochromatin formation by maintaining overall chromatin structure and, in particular, that of constitutive heterochromatin by stabilizing histone methylation (By similarity). Recruits and targets histone methyltransferases KMT5B and KMT5C, leading to epigenetic transcriptional repression (By similarity). Controls histone H4 'Lys-20' trimethylation (By similarity). Probably acts as a transcription repressor by recruiting chromatin-modifying enzymes to promoters (By similarity). Potent inhibitor of E2F-mediated trans-activation (PubMed:8319904). May act as a tumor suppressor.

Chromosomal Localization:

20q11.23

Accession Number:

UniProt Accession Number:

P28749

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

CSIYI-pS-PHKN

Native Sequence:

Calculated Isoelectric Point:

Molecular Weight:

1100

Last Updated:

11/18/2022

Links

Characterization Data

SOPs

No SOPs available.

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