详细说明
Species Reactivity
Human
Specificity
Detects human B7‑1/CD80 in ELISAs. In sandwich immunoassays, no cross-reactivity or interference with recombinant human (rh) B7-2, recombinant mouse (rm) B7-1, rmB7-2, rhB7-H1, rhB7-H2, rhB7-H3 or rmPD-L2 is observed.
Source
Monoclonal Mouse IgG 1 Clone # 37711
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human B7‑1/CD80
Val35-Asn242 (predicted)
Accession # P33681Formulation
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
ConcentrationSample
Flow Cytometry
2.5 µg/10 6 cells
See below
Immunohistochemistry
8-25 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Human B7-1/CD80 Sandwich Immunoassay
Reagent
ELISA Capture (Matched Antibody Pair)
2-8 µg/mL
Human B7‑1/CD80 Antibody (Catalog # )
ELISA Detection (Matched Antibody Pair)
0.5-2.0 µg/mL
Human B7‑1/CD80 Biotinylated Antibody (Catalog # )
ELISA Standard
Recombinant Human B7-1/CD80 Fc Chimera His-tag Protein, CF (Catalog # )
Neutralization
Measured by its ability to neutralize B7‑1/CD80 and PHA-induced IL‑2 secretion in the Jurkat human acute T cell leukemia cell line. The Neutralization Dose (ND 50) is typically 0.2-1 µg/mL in the presence of 1 µg/mL Recombinant Human B7‑1/CD80 Fc Chimera and 10 µg/mL PHA.
Please Note: Optimal dilutions should be determined by each laboratory for each application. are available in the Technical Information section on our website.
Data Examples
Flow Cytometry | Detection of B7‑1/CD80 in Raji Human Cell Line by Flow Cytometry. Raji human Burkitt's lymphoma cell line was stained with Mouse Anti-Human B7‑1/CD80 Monoclonal Antibody (Catalog # MAB140, filled histogram) or isotype control antibody (Catalog # , open histogram), followed by Fluorescein-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # ). |
Neutralization | IL‑2 secretion Induced by B7‑1/CD80 and Neutralization by Human B7‑1/CD80 Antibody. Recombinant Human B7‑1/CD80 Fc Chimera (Catalog # ) 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 # ). IL‑2 secretion elicited by Recombinant Human B7‑1/CD80 Fc Chimera (1 µg/mL) and PHA (10 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human B7‑1/CD80 Monoclonal Antibody (Catalog # MAB140). The ND50 is typically 0.2-1 µg/mL. |
Immunohistochemistry | B7‑1/CD80 in Human Tonsil. B7‑1/CD80 was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Human B7‑1/CD80 Monoclonal Antibody (Catalog # MAB140) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # ) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for . |
Preparation and Storage
Reconstitution
Reconstitute at 0.5 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 co-stimulatory 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. Human B7-1 is a 288 amino acid (aa) protein containing a 34 aa signal peptide, a 208 aa extracellular domain, a 21 aa transmembrane domain, and a 25 aa cytoplasmic domain. Human B7-1 and B7-2 share 26% aa sequenceidentity. Human and mouse B7-1 share 44% aa sequenceidentity. 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:
Azuma, M. et al. (1993) Nature 366:76.
Freeman, G.J. et al. (1993) Science 262:909.
Freeman, G. et al. (1991) J. Exp. Med. 174:625.
Selvakumar, A. et al. (1993) Immunogenetics 38:292.
Chen, C. et al. (1994) J. Immunol. 152:4929.
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