Overview

  • Product nameAnti-Rb antibody
    See all Rb primary antibodies
  • Description
    Rabbit polyclonal to Rb
  • Tested applicationsSuitable for: IHC-P, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Dog, Chimpanzee
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 800 - 900 of Human Rb.

    (Peptide available as ab94885.)

  • Positive control
    • WB: HeLa, Jurkat, Y79 and WERI whole cell lysates, mouse lung, mouse kidney, rat kidney tissue lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab85607 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
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 106 kDa (predicted molecular weight: 106 kDa).

Target

  • FunctionKey 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 specificityExpressed in the retina.
  • Involvement in diseaseChildhood cancer retinoblastoma
    Bladder cancer
    Osteogenic sarcoma
  • Sequence similaritiesBelongs to the retinoblastoma protein (RB) family.
  • DomainThe 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 localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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_HUMAN antibody
    • RB1 antibody
    • RB1 gene antibody
    • Retinoblastoma 1 antibody
    • Retinoblastoma suspectibility protein antibody
    • Retinoblastoma-associated protein antibody
    see all

Anti-Rb antibody images

  • All lanes : Anti-Rb antibody (ab85607) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : WERI (Human Retinoblastoma) Whole Cell Lysate
    Lane 4 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 106 kDa
    Observed band size : 106 kDa
    Additional bands at : 50 kDa,68 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
  • All lanes : Anti-Rb antibody (ab85607) at 1 µg/ml

    Lane 1 : Lung (Mouse) Tissue Lysate
    Lane 2 : Kidney (Mouse) Tissue Lysate
    Lane 3 : Kidney (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 106 kDa
    Observed band size : 106 kDa
    Additional bands at : 35 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 106 kDa


    Exposure time : 10 seconds
  • ICC/IF image of ab85607 stained HepG2 cells. The cells were 5% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85607, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa cells at 5µg/ml.
  • IHC image of Ab85607 staining in Human Cervical Carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with Ab85607, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References for Anti-Rb antibody (ab85607)

ab85607 has not yet been referenced specifically in any publications.

Product Wall

Thank you for your inquiry.

I am happy to confirm that ab85607 is already tested and guaranteed for WB with rat and mouse samples. Your customer will be able to take advantage of our Abpromise in the unlikely case the antibody proves difficul...

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Thank you for your reply and for kindly providing these details.

Regrettably, I am sorry ab23474 Anti-TATA binding protein TBP antibodyis not covered by our guarantee for rat samples. I appreciate your concerns and it is regrettable the resu...

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Thank you for contacting Abcam and for your help in finding the associated proteins on the phone.

I have had the chance to run the alignments of these proteins and have found that the homology between them is extremely poor. As such I do not...

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Thanks for the information.

I have initiated the replacement order. Please let me know if you do not receive it shortly.

I'm sorry this vial of ab85607 is giving you trouble. Thanks for taking the time to submit your completed questionnaire and for including the ICC and western blot images.

It does seem that the anti-Rb antibody has an unexpected staining patte...

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