Recombinant
RabMAb

Recombinant Anti-Rb (phospho T252) antibody [EPR17733] (ab184797)

Overview

  • Product name

    Anti-Rb (phospho T252) antibody [EPR17733]
    See all Rb primary antibodies
  • Description

    Rabbit monoclonal [EPR17733] to Rb (phospho T252)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Dot blot, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Rb aa 200-300 (phospho T252). The exact sequence is proprietary.
    Database link: P06400

  • Positive control

    • WB: K562 and MCF7 whole cell lysates. IHC-P: Human colon and squamous cell carcinoma of cervix tissues.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab184797 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Dot blot 1/1000.
WB 1/1000. Detects a band of approximately 106 kDa (predicted molecular weight: 106 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • 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). 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.
  • Tissue specificity

    Expressed in the retina.
  • Involvement in disease

    Childhood cancer retinoblastoma
    Bladder cancer
    Osteogenic sarcoma
  • Sequence similarities

    Belongs to the retinoblastoma protein (RB) family.
  • Domain

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

    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.
    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.
    Acetylation at Lys-873 and Lys-874 regulates subcellular localization, at least during keratinocytes differentiation.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

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

Images

  • All lanes : Anti-Rb (phospho T252) antibody [EPR17733] (ab184797) at 1/1000 dilution

    Lane 1 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 2 : K562 whole cell lysate serum starved for 3 days

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 106 kDa
    Observed band size: 106 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 2% BSA /TBST.

  • All lanes : Anti-Rb (phospho T252) antibody [EPR17733] (ab184797) at 1/1000 dilution

    Lane 1 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 2 : MCF7 whole cell lysate serum starved for 3 days

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 106 kDa
    Observed band size: 106 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 2% BSA /TBST.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Rb (phospho T252) with ab184797 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear staining on sporadic epithelium cells of Human colon is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Human squamous cell carcinoma of cervix tissue labeling Rb (phospho T252) with ab184797 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear staining on tumor cells of Human squamous cell carcinoma of cervix.is observed.

    Counter stained with Hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Dot blot analysis of Rb (phospho T252) phospho peptide (Lane 1) and Rb Non-phospho peptide (Lane 2) labeled using ab184797 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody (ab97051) at 1/1000 dilution.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

References

ab184797 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton X-100
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Jul 03 2018

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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