Recombinant
RabMAb

Recombinant Anti-B7H4 antibody [EPR20236] (ab209242)

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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

    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 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.
  • Information by UniProt

  • Database links

    • 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.

    References

    This product has been referenced in:

    • 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). Read more (PubMed: 31223291) »
    • 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). Read more (PubMed: 29436630) »
    See all 3 Publications for this product

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

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