Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
  • Flow Cytometry - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
  • Western blot - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
  • Western blot - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
  • Western blot - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
  • Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR1157(2)] to CD80 - Low endotoxin, Azide free
  • Suitable for: Flow Cyt, WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free
    See all CD80 primary antibodies

  • Description

    Rabbit monoclonal [EPR1157(2)] to CD80 - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human CD80 aa 250 to the C-terminus.

  • Positive control

    • WB: Raji and Ramos cell lysates. IHC-P: Human tonsil tissue. Flow Cyt: Raji cells.

  • General notes

    ab224797 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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

Applications

Our Abpromise guarantee covers the use of ab224797 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
Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 60 kDa (predicted molecular weight: 33 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for ICC/IF.

    • Target

    • Function

      Involved in the costimulatory signal essential for T-lymphocyte activation. T-cell proliferation and cytokine production is induced by the binding of CD28 or CTLA-4 to this receptor.

    • Tissue specificity

      Expressed on activated B-cells, macrophages and dendritic cells.

    • Sequence similarities

      Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
      Contains 1 Ig-like V-type (immunoglobulin-like) domain.

    • Cellular localization

      Membrane.
    • Information by UniProt

    • Database links

      • Alternative names

        • Activation B7-1 antigen antibody
        • B lymphocyte activation antigen B7 antibody
        • B7 antibody
        • B7-1 antibody
        • B7-1 antigen antibody
        • B7.1 antibody
        • BB1 antibody
        • CD28 antigen ligand 1 antibody
        • CD28LG antibody
        • CD28LG1 antibody
        • CD80 antibody
        • CD80 antigen (CD28 antigen ligand 1, B7-1 antigen) antibody
        • CD80 antigen antibody
        • CD80 molecule antibody
        • CD80_HUMAN antibody
        • Costimulatory factor CD80 antibody
        • costimulatory molecule variant IgV-CD80 antibody
        • CTLA-4 counter-receptor B7.1 antibody
        • LAB7 antibody
        • T-lymphocyte activation antigen CD80 antibody
        see all

      Images

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)

        This IHC data was generated using the same anti-CD80 antibody clone [EPR1157(2)] in a different buffer formulation (cat# ab134120).

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD80 with ab134120 at a dilution of 1/1000. Heat mediated antigen retrieval was performed using Trs/EDTA buffer, pH9 (ab93684). ImmunoHistoProbe one step HRP Polymer was used. Counter stained with hematoxylin.

        The image shows cytoplasmic and membrane staining.

      • Flow Cytometry - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
        Flow Cytometry - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)

        Flow Cytometry analysis of Raji (human Burkitt's lymphoma B lymphocyte) cells labeling CD80 with purified ab134120 at 1:190 dilution (10.33 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224797)

      • Western blot - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
        Western blot - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
        All lanes : Anti-CD80 antibody [EPR1157(2)] (ab134120) at 1/1000 dilution

        Lane 1 : Wild-type Raji cell lysate
        Lane 2 : CD80 knockout Raji cell lysate
        Lane 3 : Ramos cell lysate

        Lysates/proteins at 20 µg per lane.

        Performed under reducing conditions.

        Predicted band size: 33 kDa
        Observed band size: 55 kDa
        why is the actual band size different from the predicted?



        This data was developed using the same antibody clone in a different buffer formulation (ab134120).

        Lanes 1 - 3: Merged signal (red and green). Green - ab134120 observed at 55 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

        ab134120 was shown to react with CD80 in wild-type Raji cells in western blot with loss of signal observed in CD80 knockout sample. Wild-type and CD80 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab134120 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

      • Western blot - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
        Western blot - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
        All lanes : Anti-CD80 antibody [EPR1157(2)] (ab134120) at 1/1000 dilution (Unpurified)

        Lane 1 : Raji cell lysate
        Lane 2 : Ramos cell lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

        Predicted band size: 33 kDa
        Observed band size: 60 kDa why is the actual band size different from the predicted?



        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224797)

      • Western blot - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
        Western blot - Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
        All lanes : Anti-CD80 antibody [EPR1157(2)] (ab134120) at 1/1000 dilution (unpurified)

        Lane 1 : Raji cell lysate
        Lane 2 : Ramos cell lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

        Predicted band size: 33 kDa
        Observed band size: 60 kDa why is the actual band size different from the predicted?



        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224797)

      • Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
        Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)

      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 (3)

      Publishing research using ab224797? Please let us know so that we can cite the reference in this datasheet.

      ab224797 has been referenced in 3 publications.

      • Müller A  et al. Towards non-invasive imaging of vulnerable atherosclerotic plaques by targeting co-stimulatory molecules. Int J Cardiol 174:503-15 (2014). IHC ; Human . PubMed: 24834996
      • Kolyva AS  et al. The role of obesity in the immune response during sepsis. Nutr Diabetes 4:e137 (2014). IHC-P ; Human . PubMed: 25244356
      • Jia R  et al. The splicing factor hnRNP C regulates expression of co-stimulatory molecules CD80 and CD40 in dendritic cells. Immunol Lett 153:27-32 (2013). PubMed: 23831410

      Customer reviews and Q&As

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