Mouse B7-1/CD80 Antibody

(3 citations)   
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse B7‑1/CD80 in ELISAs and Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse B7‑1/CD80
    Asp37-Lys245
    Accession # Q00609
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
    • Mouse B7-1/CD80 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Mouse B7‑1/CD80 Antibody (Catalog # AF740)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Mouse B7‑1/CD80 Biotinylated Antibody (Catalog # BAF740)
  • ELISA Standard
     
    Recombinant Mouse B7-1/CD80 Fc Chimera His-tag Protein, CF (Catalog # 740-B1)
  • Neutralization
    Measured by its ability to neutralize B7‑1/CD80-induced IL‑2 secretion in the Jurkat human acute T cell leukemia cell line. The Neutralization Dose (ND50) is typically 0.15-0.6 µg/mL in the presence of 0.1 µg/mL Recombinant Mouse B7‑1/CD80 Fc Chimera and 10 µg/mL PHA.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Mouse B7‑1/CD80 by Western Blot. Western blot shows lysates of C2C12 mouse myoblast cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7‑1/CD80 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of B7‑1/CD80 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes either treated with 200 ng/mL LPS (filled histogram) or unstimulated (open histogram) were stained with Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

B7‑1/CD80 in Mouse Splenocytes.
B7‑1/CD80 was detected in immersion fixed mouse splenocytes using Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

IL‑2 secretion Induced by
B7‑1/CD80 and Neutralization by Mouse
B7‑1/CD80 Antibody.
Recombinant Mouse
B7‑1/CD80 Fc Chimera (Catalog # 740-B1) co-stimulates IL‑2 secretion in the Jurkat human acute T cell leukemia cell line in the presence of PHA in a dose-dependent manner (orange line), as measured by the Human IL‑2 Quantikine ELISA Kit (Catalog # D2050). IL‑2 secretion elicited by Recombinant Mouse
B7‑1/CD80 Fc Chimera (0.1 µg/mL) and PHA (10 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740). The ND50 is typically
0.15‑0.6 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: B7-1/CD80

B7-1 and B7-2, together with their receptors CD28 and CTLA-4, constitute one of the dominant costimulatory pathways that regulate T- and B-cell responses. Although both CTLA-4 and CD28 can bind to the same ligands, CTLA-4 binds to B7-1 and B7-2 with a 20‑100 fold higher affinity than CD28 and is involved in the
down‑regulation of the immune response. B7-1 is expressed on activated B cells, activated T cells, and macrophages. B7-2 is constitutively expressed on interdigitating dendritic cells, Langerhans cells, peripheral blood dendritic cells, memory B cells, and germinal center B cells. Additionally, B7-2 is expressed at low levels on monocytes and can be up-regulated through interferon gamma. B7-1 and B7-2 are both members of the immunoglobulin superfamily. Mouse B7-1 is a 306 amino acid (aa) protein containing a putative 37 aa signal peptide, a 190 aa extracellular domain, a 22 aa transmembrane domain, and a 38 aa cytoplasmic domain. Mouse B7-1 and B7-2 share 28% amino acid identity. Mouse and human B7-1 share 44% amino acid identity. However, it has been observed that both human and mouse
B7‑1 and B7‑2 can bind to either human or mouse CD28 and CTLA-4, suggesting that there are conserved amino acids which form the B7-1/B7-2/CD28/CTLA-4 critical binding sites.

  • References:
    1. Azuma, M. et al. (1993) Nature 366:76.
    2. Freeman, G.J. et al. (1993) Science 262:909.
    3. Freeman, G. et al. (1991) J. Exp. Med. 174:625.
    4. Selvakumar, A. et al. (1993) Immunogenetics 38:292.
    5. Chen, C. et al. (1994) J. Immunol. 152:4929.
    6. Freeman, G.J. et al. (1993) J. Exp. Med. 178:2185.
  • Entrez Gene IDs:
    941 (Human); 12519 (Mouse); 25408 (Rat); 102140078 (Cynomolgus Monkey)
  • Alternate Names:
    Activation B7-1 antigen; B7; B71; B7-1; BB1; CD28LG1; CD28LGB7-1 antigen); CD80 antigen; CD80 molecule; CD80; costimulatory factor CD80; costimulatory molecule variant IgV-CD80; CTLA-4 counter-receptor B7.1; T-lymphocyte activation antigen CD80
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. Aging is associated with increased regulatory T-cell function.
    Authors: Garg S, Delaney C, Toubai T, Ghosh A, Reddy P, Banerjee R, Yung R
    Aging Cell, 2014;13(3):441-8.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  2. Antigen-loaded ER microsomes from APC induce potent immune responses against viral infection.
    Authors: Sofra V, Mansour S, Liu M, Gao B, Primpidou E, Wang P, Li S
    Eur. J. Immunol., 2009;39(1):85-95.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  3. Suppression of activation and induction of apoptosis in RAW264.7 cells by amniotic membrane extract.
    Authors: He H, Li W, Chen SY, Zhang S, Chen YT, Hayashida Y, Zhu YT, Tseng SC
    Invest. Ophthalmol. Vis. Sci., 2008;49(10):4468-75.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
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