Recombinant
RabMAb

Recombinant Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

Rabbit recombinant monoclonal Rb (phospho S780) antibody [EPR182(N)]. Validated in WB, IHC, Dot and tested in Human. Cited in 5 publication(s).

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Overview

  • Product name

    Anti-Rb (phospho S780) antibody [EPR182(N)]
    See all Rb primary antibodies

  • Description

    Rabbit monoclonal [EPR182(N)] to Rb (phospho S780)

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Dot blot, IHC-P, WBmore details
    Unsuitable for: Flow Cyt,ICC or IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Rb aa 750-850 (phospho S780) (Cysteine residue). The exact sequence is proprietary.
    Database link: P06400

  • Positive control

    • Jurkat cell lysate; Human thyroid gland carcinoma and testis tissues.

  • General notes

    A trial size is available to purchase for this antibody.

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab173289 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.
IHC-P 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/10000. Predicted molecular weight: 106 kDa.
  • Application notes
    Is unsuitable for Flow Cyt,ICC or IP.

    • 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

      • Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        All lanes : Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289) at 1/50000 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
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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


        Exposure time: 1 minute


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

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

        Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Rb (phospho S780) with ab173289 at 1/100 dilution,followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

         

        Nucleus staining on normal human colon.

         

        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 citrate buffer pH 6 before commencing with IHC staining protocol.

      • Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        Lane 1 : Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289) at 1/20000 dilution
        Lane 2 : Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

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

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

        Predicted band size: 106 kDa



        Blocking/Dilution buffer:5% NFDM/TBST

        Exposure time:1 minute

      • Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        All lanes : Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289) at 1/5000 dilution

        Lane 1 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
        Lane 2 : Calyculin A treated K562(Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate

        Lysates/proteins at 10 µg per lane.

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

        Predicted band size: 106 kDa



        Blocking/Dilution buffer:NFDM/TBST

        Ab173289: anti- Rb (phospho S780);

        Ab184796: anti- Rb (phospho S807);

        Ab181616: anti-Rb

         

      • Dot Blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        Dot Blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

        Dot blot analysis of Rb (phospho S780) peptide (Lane 1),Rb(non-phospho) peptide (Lane 2) with ab173289 at a dilution of 1/1000.ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/1000.

        Blocking and dilution buffer:5% NFDM/TBST.

        Exposure time:3 minutes

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

        Immunohistochemical analysis of paraffin-embedded Lung squamous cell carcinoma labeling Rb (phospho S780) with ab173289 at 1/100 dilution,followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

         

        Nucleus staining on tumor cells of lung squamous cell carcinoma.

        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 citrate buffer pH 6 before commencing with IHC staining protocol.

      • Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        All lanes : Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289) at 1/1000 dilution

        Lane 1 : Jurkat cell lysate
        Lane 2 : Jurkat cell lysate treated with Lambda Phosphatase

        Lysates/proteins at 10 µg per lane.

        Predicted band size: 106 kDa

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

        Immunohistochemical analysis of paraffin-embedded Human thyroid gland carcinoma tissue labeling Rb with ab173289 at 1/50 dilution.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

        Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Rb with ab173289 at 1/50 dilution.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      References

      This product has been referenced in:

      • Chen JJ  et al. Antipsychotic agent pimozide promotes reversible proliferative suppression by inducing cellular quiescence in liver cancer. Oncol Rep 42:1101-1109 (2019). Read more (PubMed: 31322218) »
      • Sun Y  et al. Potent anti-tumor efficacy of palbociclib in treatment-naïve H3.3K27M-mutant diffuse intrinsic pontine glioma. EBioMedicine 43:171-179 (2019). Read more (PubMed: 31060906) »
      See all 8 Publications for this product

      Publishing research using ab173289? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

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      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Rb (phospho S780) antibody [EPR182(N)]

       Excellent
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Human Tissue sections (human primary incisor tooth germ)
      Antigen retrieval step
      Heat mediated - Buffer/Enzyme Used: citrate buffer
      Permeabilization
      No
      Specification
      human primary incisor tooth germ
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
      Fixative
      Paraformaldehyde
      Read More

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      Verified customer

      Submitted Dec 25 2019

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