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RecombinantRabMAb

Recombinant Anti-B7H4 antibody [EPR20236] (ab209242)

  • Datasheet
  • Certificate of Compliance
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (ab209242)
  • Immunocytochemistry/ Immunofluorescence - Anti-B7H4 antibody [EPR20236] (ab209242)
  • Western blot - Anti-B7H4 antibody [EPR20236] (ab209242)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (ab209242)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (ab209242)
  • Anti-B7H4 antibody [EPR20236] (ab209242)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20236] to B7H4
  • Suitable for: ICC/IF, IHC-P, WB
  • Reacts with: Human

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Overview

  • Product name

    Anti-B7H4 antibody [EPR20236]
    See all B7H4 primary antibodies
  • Description

    Rabbit monoclonal [EPR20236] to B7H4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human B7H4 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q7Z7D3

  • Positive control

    • WB: SK-BR-3 and T-47D whole cell lysates; Human breast cancer lysate. IHC-P: Human breast, breast cancer and kidney tissues. ICC/IF: SK-BR-3 cells.
  • General notes

    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.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20236
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Other
    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Cancer
    • Cell cycle
    • Cell cycle inhibitors
    • Other
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells

Associated products

  • Alternative Versions

    • Anti-B7H4 antibody [EPR20236] - Low endotoxin, Azide free (ab222927)
    • Alexa Fluor® 488 Anti-B7H4 antibody [EPR20236] (ab225487)
    • Alexa Fluor® 647 Anti-B7H4 antibody [EPR20236] (ab225488)
    • Anti-B7H4 antibody [EPR20236] - BSA and Azide free (ab271987)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human B7H4 protein (Biotin) (ab198650)

Applications

Our Abpromise guarantee covers the use of ab209242 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.
IHC-P 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 65, 45, 28, 18 kDa (predicted molecular weight: 31 kDa).

Target

  • Function

    Negatively regulates T-cell-mediated immune response by inhibiting T-cell activation, proliferation, cytokine production and development of cytotoxicity. When expressed on the cell surface of tumor macrophages, plays an important role, together with regulatory T-cells (Treg), in the suppression of tumor-associated antigen-specific T-cell immunity. Involved in promoting epithelial cell transformation.
  • Tissue specificity

    Overexpressed in breast, ovarian, endometrial, renal cell (RCC) and non-small-cell lung cancers (NSCLC). Expressed on activated T- and B-cells, monocytes and dendritic cells, but not expressed in most normal tissues (at protein level). Widely expressed, including in kidney, liver, lung, ovary, placenta, spleen and testis.
  • Sequence similarities

    Belongs to the immunoglobulin superfamily. BTN/MOG family.
    Contains 2 Ig-like V-type (immunoglobulin-like) domains.
  • Post-translational
    modifications

    N-glycosylated.
  • Cellular localization

    Cell membrane. Expressed at the cell surface. A soluble form has also been detected.
  • Target information above from: UniProt accession Q7Z7D3 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 79679 Human
    • Omim: 608162 Human
    • SwissProt: Q7Z7D3 Human
    • Unigene: 546434 Human
    • Alternative names

      • B7 family member, H4 antibody
      • B7 H4 antibody
      • B7 homolog 4 antibody
      • B7 superfamily member 1 antibody
      • B7 superfamily, member 1 antibody
      • B7-H4 antibody
      • B7h.5 antibody
      • B7h4 antibody
      • B7S1 antibody
      • B7x antibody
      • BC032925 antibody
      • Immune costimulatory protein B7-H4 antibody
      • Immune costimulatory protein B7H4 antibody
      • MGC41287 antibody
      • PRO1291 antibody
      • Protein B7S1 antibody
      • RP11 229A19.4 antibody
      • T cell costimulatory molecule B7x antibody
      • T-cell costimulatory molecule B7x antibody
      • V set domain-containing T cell activation inhibitor 1 antibody
      • V-set domain-containing T-cell activation inhibitor 1 antibody
      • VCTN1 antibody
      • Vtcn1 antibody
      • VTCN1_HUMAN antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (ab209242)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (ab209242)

      Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling B7H4 with ab209242 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous and weak cytoplasmic staining on tumor cells of human breast cancer is observed [PMID:15756008]. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

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

    • Immunocytochemistry/ Immunofluorescence - Anti-B7H4 antibody [EPR20236] (ab209242)
      Immunocytochemistry/ Immunofluorescence - Anti-B7H4 antibody [EPR20236] (ab209242)

      Immunofluorescent analysis of 100% methanol-fixed SK-BR-3 (Human mammary gland adenocarcinoma cell line) cells labeling B7H4 with ab209242 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on SK-BR-3 cell line. 

      The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    • Western blot - Anti-B7H4 antibody [EPR20236] (ab209242)
      Western blot - Anti-B7H4 antibody [EPR20236] (ab209242)
      All lanes : Anti-B7H4 antibody [EPR20236] (ab209242) at 1/1000 dilution

      Lane 1 : SK-BR-3 (Human mammary gland adenocarcinoma cell line) whole cell lysate
      Lane 2 : T-47D (Human ductal breast epithelial tumor cell line) whole cell lysate
      Lane 3 : Human breast cancer lysate
      Lane 4 : HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate
      Lane 5 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 31 kDa
      Observed band size: 18,28,45,65 kDa
      why is the actual band size different from the predicted?


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      B7H4 is N-glycosylated protein. The level of B7H4 glycosylation varies between different cells and tissues. The bands at 65kDa and 45kDa represent glycosylated B7H4. The bands at 28kDa and 18kDa represent de-glycosylated B7H4. HT-29 and HeLa cell lines are negative controls. (PMID: 15878339).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (ab209242)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (ab209242)

      Immunohistochemical analysis of paraffin-embedded human breast tissue labeling B7H4 with ab209242 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Predominantly apical, luminal membranous staining on human breast epithelium is observed [PMID: 15756008]. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (ab209242)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (ab209242)

      Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling B7H4 with ab209242 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous staining on distal convoluted renal tubules of human kidney is observed [PMID: 16798883]. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

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

    • Anti-B7H4 antibody [EPR20236] (ab209242)
      Anti-B7H4 antibody [EPR20236] (ab209242)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (5)

    Publishing research using ab209242? Please let us know so that we can cite the reference in this datasheet.

    ab209242 has been referenced in 5 publications.

    • Hu X  et al. The anti-B7-H4 checkpoint synergizes trastuzumab treatment to promote phagocytosis and eradicate breast cancer. Neoplasia 22:539-553 (2020). PubMed: 32966956
    • Cao H  et al. Clinical value of detecting IQGAP3, B7-H4 and cyclooxygenase-2 in the diagnosis and prognostic evaluation of colorectal cancer. Cancer Cell Int 19:163 (2019). PubMed: 31223291
    • Yuan X  et al. Blockade of Immune-Checkpoint B7-H4 and Lysine Demethylase 5B in Esophageal Squamous Cell Carcinoma Confers Protective Immunity against P. gingivalis Infection. Cancer Immunol Res 7:1440-1456 (2019). PubMed: 31350278
    • Zhou D  et al. Silencing of B7-H4 suppresses the tumorigenicity of the MGC-803 human gastric cancer cell line and promotes cell apoptosis via the mitochondrial signaling pathway. Int J Oncol 52:1267-1276 (2018). PubMed: 29436630
    • Kang FB  et al. B7-H4 overexpression is essential for early hepatocellular carcinoma progression and recurrence. Oncotarget 8:80878-80888 (2017). IHC-P ; Human . PubMed: 29113351

    Customer reviews and Q&As

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