Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-B7H4 antibody [EPR20236-9] - BSA and Azide free (ab269460)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20236-9] to B7H4 - BSA and Azide free
  • Suitable for: WB, ICC/IF, Flow Cyt, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-B7H4 antibody [EPR20236-9] - BSA and Azide free
    See all B7H4 primary antibodies

  • Description

    Rabbit monoclonal [EPR20236-9] to B7H4 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IPmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: SK-BR-3, T47D and Mouse uterus lysates. ICC/IF: HEK-293T cell line transfected with a HA-tagged B7-H4 expression vector. Flow Cyt: 293T (human embryonic kidney epithelial cell) transfected with myc-tagged B7-H4 expression vector. IP: SK-BR-3 cells.

  • General notes

    ab269460 is the carrier-free version of ab269455. 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

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

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

Images

  • Immunoprecipitation - Anti-B7H4 antibody [EPR20236-9] - BSA and Azide free (ab269460)
    Immunoprecipitation - Anti-B7H4 antibody [EPR20236-9] - BSA and Azide free (ab269460)

    B7H4 was immunoprecipitated from 0.35 mg SK-BR-3 (Human breast adenocarcinoma epithelial cell) whole cell lysate with ab269455 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab269455 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

    Lane 1: SK-BR-3 whole cell lysate 10ug.

    Lane 2: ab269455 IP in SK-BR-3 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab269455 in SK-BR-3 whole cell lysate.

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

    Exposure time: 30 seconds.

    B7H4 is N-glycosylated protein. The level of B7H4 glycosylation varies between different cells and tissues. The band at 65kDa represents glycosylated B7H4. The band at 28kDa represents non-glycosylated B7H4 (PMID: 15878339).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269455).

  • Flow Cytometry - Anti-B7H4 antibody [EPR20236-9] - BSA and Azide free (ab269460)
    Flow Cytometry - Anti-B7H4 antibody [EPR20236-9] - BSA and Azide free (ab269460)

    Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized HEK-293T (human embryonic kidney epithelial cell) transfected with myc-tagged B7-H4 expression vector cells labelling B7H4 with ab269455 at 1/60 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left). PMID: 23722540 and PMID: 27632942 showed that cell surface expression of B7-H4 hard to detected in vitro cultured cell lines, and all the commercial B7H4 antibodies that recommend FC as an application use the overexpression system. Our RabMab also could recognise overexpressed B7H4.The overexpression plasmid contains the full length aa1-282 of human B7H4(Q7Z7D3) with HA tag in pcDNA3.1(+) vector.

    Cells were surface stained with rabbit IgG (Left) or ab269455 (Right). Then fixed with 2% PFA followed by intracellularly stained with anti-HA tag conjugated to Alexa Fluor® 647.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269455).

  • Immunocytochemistry/ Immunofluorescence - Anti-B7H4 antibody [EPR20236-9] - BSA and Azide free (ab269460)
    Immunocytochemistry/ Immunofluorescence - Anti-B7H4 antibody [EPR20236-9] - BSA and Azide free (ab269460)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human embryonic kidney epithelial cell) cells labelling B7H4 with ab269455 at 1/500 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and membranous staining in HEK-293T cell line transfected with a HA-tagged B7-H4 expression vector. Alexa Fluor® 647 anti-HA.11 Epitope Tag Antibody was used at 200 dilution to detecte the overexpressed protein (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 AlexaFluo®488 Goat anti-Rabbit secondary at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269455).

  • Anti-B7H4 antibody [EPR20236-9] - BSA and Azide free (ab269460)
    Anti-B7H4 antibody [EPR20236-9] - BSA and Azide free (ab269460)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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