Mouse B7-1/CD80 Antibody

R&D Systems | Catalog # AF740

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse

Applications

Validated:

Western Blot, ELISA Capture (Matched Antibody Pair), Neutralization, Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all B7-1/CD80 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse B7‑1/CD80
Asp37-Lys245
Accession # Q00609

Specificity

Detects mouse B7‑1/CD80 in ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Mouse B7-1/CD80 Antibody

Detection of Mouse B7-1/CD80 antibody by Western Blot.

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 antibody in Mouse Splenocytes antibody by Flow Cytometry.

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.

Detection of B7‑1/CD80 in Mouse Splenocytes by Flow Cytometry

Mouse B6 splenocytes treated with 200 ng/mL LPS for 48 hr were stained with (A) Goat Anti-Mouse B7-1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740) or (B) Goat IgG control antibody (AB-108-C) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (F0107) and Rat Anti-Mouse B220/CD45R Alexa Fluor® 488-conjugated Monoclonal Antibody (FAB1217G). View our protocol for Staining Membrane-associated Proteins.
B7-1/CD80 antibody in Mouse Splenocytes by Immunocytochemistry (ICC).

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.

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.
Mouse B7-1 / CD80 ELISA Standard Curve

Mouse B7-1 / CD80 ELISA Standard Curve

Recombinant Mouse B7‑1/CD80 Fc Chimera (Catalog # 740-B1) was serially diluted and captured by Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)
Mouse B7-1 / CD80 ELISA Standard Curve

Mouse B7-1 / CD80 ELISA Standard Curve

Recombinant Mouse B7‑1/CD80 Fc Chimera (Catalog # 740-B1) was serially diluted and captured by Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)
Detection of B7-1/CD80 by Flow Cytometry

Detection of B7-1/CD80 by Flow Cytometry

Single-cell sequencing reveals an inhibitory role of neutrophils in tumor immune status through CD80/CD86-CTLA4 axis.A, B Interactions between subtypes of immune cells were analyzed. Functional phenotypes and predicted interactions of myeloid cells and T cells (A), and ligand-receptor interactions in immune checkpoints between immune cells are shown (B). C Myeloid cells were divided into 2 clusters (N2 and non-N2) using a UMAP plot. D The expression level of CD80, CD86, CCL5 and CSF1R in N2 and non-N2 cluster is shown. E Murine T cells, T cells pre-stimulated with neutrophils, T cells pre-stimulated with neutrophils in the presence of CD80 + CD86 neutralizing antibodies (anti-CD80/CD86) were pre-treated with PD-1 neutralizing antibodies and then co-cultured with MC38 cells for 6 h. Apoptosis assays were used to evaluate the proportion of apoptosis in each group. The two-tailed unpaired Student’s t test was used. The mean values are shown, and the standard deviations are displayed by the error bars. Each experiment was repeated 3 times with 3 replicates. Source data are provided as a Source Data file. Neu neutrophil. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36443332), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse B7-1/CD80 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: Mouse splenocytes treated with LPS

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed mouse splenocytes

Western Blot

1 µg/mL
Sample: C2C12 mouse myoblast cell line

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.

Mouse B7-1/CD80 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Mouse B7‑1/CD80 Biotinylated Antibody (Catalog # BAF740)
  • Standard: Recombinant Mouse B7-1/CD80 Fc Chimera His-tag Protein, CF (Catalog # 740-B1)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using AF740 in the following applications:

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Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.

Alternate Names

B71, CD80

Entrez Gene IDs

941 (Human); 12519 (Mouse); 25408 (Rat); 102140078 (Cynomolgus Monkey)

Gene Symbol

CD80

UniProt

Q00609

Additional B7-1/CD80 Products

Product Documents for Mouse B7-1/CD80 Antibody

Certificate of Analysis

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Product Specific Notices for Mouse B7-1/CD80 Antibody

For research use only

Citations for Mouse B7-1/CD80 Antibody

Customer Reviews for Mouse B7-1/CD80 Antibody (2)

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  • Mouse B7-1/CD80 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Lung tissue
    Species: Mouse
    Verified Customer | Posted 12/12/2021
    Mouse B7-1/CD80 Antibody AF740
  • Mouse B7-1/CD80 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Pancreas tissue
    Species: Mouse
    Verified Customer | Posted 12/18/2018

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