Recombinant
RabMAb

Anti-Rb (phospho S807) antibody [EPR17732] (ab184796)

Overview

  • Product name
    Anti-Rb (phospho S807) antibody [EPR17732]
    See all Rb primary antibodies
  • Description
    Rabbit monoclonal [EPR17732] to Rb (phospho S807)
  • Tested applications
    Suitable for: Dot blot, WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Rb aa 800-900 (phospho S807). The exact sequence is proprietary.
    Database link: P06400

  • Positive control
    • WB: K562, MCF7, L-929 and C6 whole cell lysates; Mouse embryo lysate. IHC-P: Human colon, Human squamous cells carcinoma of lung, mouse spleen and rat spleen tissues. ICC/IF: MCF7 cells.
  • 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 ab184796 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).
ICC/IF 1/500.
IHC-P 1/500. 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 S807) antibody [EPR17732] (ab184796) at 1/20000 dilution

    Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) serum starved for 44 h. Whole cell lysates
    Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) serum starved for 24 h. Then 10% FBS incubated for 20 h. Whole cell lysates
    Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) serum starved for 24 h. Then 10% FBS incubated for 20 h. Whole cell lysates. Then the membrane was incubated with phosphatase.

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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


    Exposure time : 10 seconds

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% tritonX-100 permeabilized MCF7 (Human breast denocarcinoma cell line) cells, Serum starved and non-starved, labeling Rb (phospho S807) with Ab184796 at 1/200 dilution followed by Goat anti-Rabbit secondary IgG AlexaFluor®488 (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing positive staining on MCF7 cells. The number of positive cells increased after treatment with FBS (fetal bovine serum) for 48 hours, then decreased after Lambda Protein Phosphatase treatment (31℃ for 2 hours ).

  • All lanes : Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/20000 dilution

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

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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


    Exposure time : 3 minutes

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/5000 dilution

    Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysates
    Lane 2 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysates treated with Lambda Phosphatase.

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG (H+L), Peroxidase conjugated at 1/1000 dilution
    Lane 2 : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution


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


    Exposure time : 30 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/1000 dilution + L-929 (Mouse connective tissue fibroblast cells) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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


    Exposure time : 3 minutes

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/2000 dilution + Mouse embryo lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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


    Exposure time : 3 minutes

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/2000 dilution + C6 (Rat glial tumor cells) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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


    Exposure time : 30 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/1000 dilution + MCF7(Human breast adenocarcinoma cell line) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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


    Exposure time : 3 minutes

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Rb (phospho S807) with ab184796 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on epithelial cells of Human colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Squamous cells carcinoma of lung tissue labeling Rb (phospho S807) with ab184796 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on squamous cells carcinoma of lung is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Rb (phospho S807) with ab184796 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling Rb (phospho S807) with ab184796 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on lymphocytes of rat spleen is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Dot blot analysis of Rb (phospho S807) peptide (Lane 1), and non-phospho peptide (Lane 2), labeled using ab184796 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution.

    Blocking/Dilution buffer: 5% NFDM/TBST.

References

This product has been referenced in:
  • Bu S  et al. Human amniotic epithelial cells inhibit growth of epithelial ovarian cancer cells via TGF-ß1-mediated cell cycle arrest. Int J Oncol 51:1405-1414 (2017). IHC-P ; Mouse . Read more (PubMed: 29048644) »

See 1 Publication for this product

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