RabMAb

Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)

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Immunocytochemistry/ Immunofluorescence - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)
  • Immunoprecipitation - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)
  • Dot Blot - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)

Key features and details

  • Rabbit monoclonal [EP821Y] to Rb (phospho T373) - BSA and Azide free
  • Suitable for: IHC-P, ICC/IF, IP, Dot blot, WB
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free
    See all Rb primary antibodies

  • Description

    Rabbit monoclonal [EP821Y] to Rb (phospho T373) - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human Rb (phospho T373). The exact sequence is proprietary.

  • General notes

    ab219158 is the carrier-free version of ab52975 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab219158 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab219158 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 at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Dot blot Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 110 kDa (predicted molecular weight: 106 kDa).
  • Application notes
    Is unsuitable for Flow Cyt.

    • 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

      • Immunocytochemistry/ Immunofluorescence - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)
        Immunocytochemistry/ Immunofluorescence - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)

        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 T373) with Ab52975 at 1/100 dilution followed by Goat anti-Rabbit secondary IgG AlexaFluor®488 (ab150077) secondary antibody at 1/1000 dilution (green).

        Confocal image showing nuclear 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 2hours ).

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52975).

      • Immunoprecipitation - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)
        Immunoprecipitation - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)
        All lanes : Anti-Rb (phospho T373) antibody [EP821Y] (ab52975) at 1/500 dilution

        Lane 1 : Jurkat (human acute T cell leukemia) starved 24h then 10% FBS incubated for 20h whole cell lysate.
        Lane 2 : Jurkat (human acute T cell leukemia ) starved 24h then 10% FBS incubated for 20h whole cell lysate.
        Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab52975 in Jurkat (human acute T cell leukemia ) starved 24h then 10% FBS incubated for 20h whole cell lysate.

        Lysates/proteins at 10 µg per lane.

        Secondary
        VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution

        Predicted band size : 106 kDa
        Additional bands at : 110 kDa.

        Exposure time : 1 second

        Blocking and diluting buffer: 5% NFDM/TBST

        The IP lysates are prepared by RIPA method. As the WB image shows, this antibody works in 1%SDS Hot Lysate buffer in WB. So there is no band in input lane.
         
        For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52975).

      • Dot Blot - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)
        Dot Blot - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)

        Dot Blot analysis of Lane 1: Rb (pT373) phospho peptide and Lane 2: Rb non-phospho peptide labeling Rb (phospho T373) with ab52975 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051 Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.

         

         

         

         

         

         

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52975).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T373) antibody [EP821Y] - BSA and Azide free (ab219158)

        ab52975, at a 1/250 dilution, staining Human Rb (Phosphorylated) in Kidney carcinoma, using Immunohistochemistry, Paraffin Embedded tissue.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52975).

        Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References (0)

      Publishing research using ab219158? Please let us know so that we can cite the reference in this datasheet.

      ab219158 has not yet been referenced specifically in any publications.

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