• Product name

    Anti-Rb antibody [E182]
    See all Rb primary antibodies
  • Description

    Rabbit monoclonal [E182] to Rb
  • Host species

  • Specificity

    The antibody detects pan Rb protein.
  • Tested applications

    Suitable for: ICC/IF, IP, WBmore details
    Unsuitable for: Flow Cyt or IHC
  • Species reactivity

    Reacts with: Human
  • Immunogen

    corresponding to Human Rb.

  • Positive control

    • WB: Jurkat, Hek293 cell lysate ICC: A431 cells
  • General notes

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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.



Our Abpromise guarantee covers the use of ab32513 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
ICC/IF 1/250 - 1/500.
IP 1/40.
WB 1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 106 kDa).
  • Application notes
    Is unsuitable for Flow Cyt or IHC.
  • 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

      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

    • 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


    • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: Rb knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: HEK293 whole cell lysate (20 µg)


      Lanes 1 - 4: Merged signal (red and green). Green - ab32513 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.


      ab32513 was shown to recognize Rb when Rb knockout samples were used, along with additional cross-reactive bands. Wild-type and Rb knockout samples were subjected to SDS-PAGE. Ab32513 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-Rb antibody [E182] (ab32513) at 1/1000 dilution

      Lane 1 : Untreated Jurkat cell lysate
      Lane 2 : Jurkat cell lysate treated with FBS
      Lane 3 : Jurkat cell lysate treated with Alkaline Phosphatase.

      Predicted band size: 106 kDa
      Observed band size: 120 kDa
      why is the actual band size different from the predicted?

    • Immunofluorescent staining of (A) non-treated and (B) Lambda Phosphatase-treated A431 cells using anti-Rb ab32513 Rabbit Monoclonal Antibody


    This product has been referenced in:

    • Richardson E  et al. Mechanism-based screen establishes signalling framework for DNA damage-associated G1 checkpoint response. PLoS One 7:e31627 (2012). ICC/IF ; Human . Read more (PubMed: 22384045) »
    • Haller F  et al. Increased KIT signalling with up-regulation of cyclin D correlates to accelerated proliferation and shorter disease-free survival in gastrointestinal stromal tumours (GISTs) with KIT exon 11 deletions. J Pathol 216:225-35 (2008). Read more (PubMed: 18729075) »
    See all 2 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A


    Thank you for contacting us.

    I have received the answers to your questions:

    1) Lambda phosphatase is usedafter fixation.
    2) Unfortunately, we do not have any product references at this time.

    3) The Lambda phosphatase treatment protocol is below:

    Materials and Reagents:

    LP Buffer
    -Combine: 2.5ml 1M TrisHCl, 0.1ml 1M MnCl2, 250ul 1M DTT, and 5mg BSA in 47ml dH2O. Adjust pH to 7.4.

    Lambda Phosphatase working solution
    -600 units LP in 1ml LP Buffer
    “Experiment” tissue/cell slide: LP blocking + IHC/ICC protocol.
    “Control” tissue/cell slide: IHC/ICC protocol


    1. Adopt tissues/cells to LP buffer:
    -Before entering the 10% goat serum blocking step in IHC or ICC, cover both the “experiment” tissue slide & “control” tissue/cell slide in LP Buffer and incubate for 45 minutes
    2. After 45 minutes LP blocking:
    -Leave “control” tissue/cell slide in LP Buffer for 1 more hour.
    -Remove LP Buffer and add LP working solution to the “experiment” tissue slide for 1 hour.
    Continue with usual IHC/ICC protocol

    LP buffer should be clear, if the solution appears brown, it is too basic.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More


    Thank you for contacting us.

    I have heard back from the lab with the following information:

    We found negative results on a cancer TMA, but no other details are available.
    The concentration varies from lot to lot, but the same batch should produce similar staining.

    I apologize that more detailed information was not available from the lab.

    As we discussed over the phone, I would suggest to include a blocking step to reduce the background.

    I hope this information is of some help to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

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