Recombinant 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
Overview
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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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20236 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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 |
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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
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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
modificationsN-glycosylated. -
Cellular localization
Cell membrane. Expressed at the cell surface. A soluble form has also been detected. - Information by UniProt
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Database links
- Entrez Gene: 79679 Human
- Omim: 608162 Human
- SwissProt: Q7Z7D3 Human
- Unigene: 546434 Human
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Alternative names
- B7 family member, H4 antibody
- B7 H4 antibody
- B7 homolog 4 antibody
see all
Images
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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.
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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.
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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 secondsBlocking/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).
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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.
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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.
Protocols
Datasheets and documents
Certificate of Compliance
References (5)
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