ELISA Development (Detection), Flow Cytometry, Functional Assay, Immunocytochemistry, Immunohistochemistry, Immunohistochemistry-Paraffin, Neutralization, T Cell Activation, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₁ Clone # 37711

View all B7-1/CD80 Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human B7‑1/CD80
Val35-Asn242 (predicted)
Accession # P33681

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Endotoxin Level

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

Scientific Data Images for Human B7-1/CD80 Antibody

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 # MAB002, open histogram), followed by Fluorescein-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0103B). View our protocol for Staining Membrane-associated Proteins.

IL‑2 secretion Induced by B7‑1/CD80 and Neutralization by Human B7‑1/CD80 Antibody.

Recombinant Human B7-1/CD80 Fc Chimera (Catalog # 140-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 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.

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 # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

B7-1/CD80 in Human Tonsil Using Dual RNAscope^®ISH and IHC.

B7-1/CD80 mRNA was detected in formalin-fixed paraffin-embedded tissue sections of human tonsil probed with ACD RNAScope^®Probe (Catalog # 42471) and stained using ACD RNAscope^®2.5 HD Duplex Detection Reagents (red, Catalog # 322500). Adjacent tissue section was processed for immunohistochemistry using R&D Systems Mouse Anti-Human B7-1/CD80 Monoclonal Antibody (Catalog# MAB140) at 5ug/mL with overnight incubation at 4 °C followed by incubation with the Anti-Mouse IgG VisUCyte HRP Polymer Antibody (R&D Systems, Catalog # VC001) and DAB chromogen (lower image, brown). Tissues were counterstained with hematoxylin (blue).

Detection of Mouse B7-1/CD80 by Immunohistochemistry

MNK inhibitors induce polarization of TAMs toward an M2 phenotype in ex vivo human PDAC slice cultures.(A) Serial sections of human PDAC tumors (n = 15) were stained for Arginase-1 and phosphorylated eIF4ES209 (p-eIF4E) by IHC. Scale bar: 200 μm. The staining positivity was analyzed by the TissueGnostics Imaging system using the HistoQuest, and the correlation analysis for Arginase-1 and p-eIF4E expression was performed using GraphPad. (B) Slice cultures (n = 1–4 replicates for each treatment group) from 6 different PDAC tumors were treated with DMSO (vehicle control) or CGP57380 for 5 days and stained for phosphorylated eIF4E (p-eIF4E), CD68, Arginase-1, CD163, MRC1, CD86, CD80, and MHCII. The percentage of positive cells, relative to the total number of nucleated cells, was analyzed by ImageJ. Scale bar: 50 μm. Data points represent individual patients. Paired t test was performed by GraphPad. *P < 0.05; **P < 0.01. (C) The relative expression of MKNK2, CD163, and MRC1 in TCGA studies and their transcript abundance from RNA-Seq data, quantified as RSEM, were downloaded from cBioPortal. Correlation analysis was performed in GraphPad to evaluate the relationship between MKNK2, CD163, and MRC1. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35380995), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Applications for Human 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.

Dual RNAscope ISH-IHC Compatible

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil tissue

Flow Cytometry

0.25 µg/10⁶ cells
Sample: Raji human Burkitt's lymphoma cell line

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil tissue

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 (ND₅₀) 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.

Human B7-1/CD80 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)

Recommended Concentration: 2-8 µg/mL
Use in combination with these reagents:

Detection Reagent: Human B7-1/CD80 Biotinylated Antibody (Catalog # BAM1402)
Standard: Recombinant Human B7-1/CD80 Fc Chimera His-tag Protein, CF (Catalog # 140-B1)

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 3.5 using MAB140 in the following applications:

Flow Cytometry (1 Review)
Immunocytochemistry/Immunofluorescence (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 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.

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.

Alternate Names

B71, CD80

Entrez Gene IDs

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

Gene Symbol

CD80

UniProt

P33681

Additional B7-1/CD80 Products

Product Documents for Human B7-1/CD80 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human B7-1/CD80 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human B7-1/CD80 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB140

Key Product Details

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Applications