Recombinant Human B7-1/CD80 Fc Chimera Protein, CF

Catalog #: 10133-B1 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
10133-B1-100
10133-B1-01M

Recombinant Human B7-1/CD80 Fc Chimera Protein Bioactivity

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Product Details

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human B7-1/CD80 Fc Chimera Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

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

Activity

Measured by its ability to induce IL-2 secretion by Jurkat human acute T cell leukemia cells. Freeman, G.J. et al. (1993) Science 262:909. The ED₅₀ for this effect is 0.025‑0.15 µg/mL in the presence of PHA.

Source

Chinese Hamster Ovary cell line, CHO-derived human B7-1/CD80 protein

Human B7-1/CD80 (Val35-Asn242) Accession # P33681	IEGRMD	Human IgG₁ (Pro100-Lys330)
N-terminus		C-terminus

Accession #

P33681

N-terminal Sequence
Analysis

Val35

Structure / Form

Disulfide-linked homodimer

Predicted Molecular Mass

50.5 kDa

SDS-PAGE

66-80 kDa, under reducing conditions

Product Datasheets

10133-B1

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

10133-B1

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 500 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it 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. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Bioactivity

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Recombinant Human B7‑1/CD80 Fc Chimera (Catalog # 10133-B1) induces IL-2 secretion by Jurkat human acute T cell leukemia cells in the presence of PHA. The ED₅₀ for this effect is 0.025-0.15 μg/mL.

SDS-PAGE

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2 μg/lane of Recombinant Human B7‑1/CD80 Fc Chimera (Catalog # 10133-B1) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 66-80 kDa and 130-160 kDa, respectively.

Surface Plasmon Resonance (SPR)

Surface plasmon resonance sensorgram of Avi-tag biotinylated human CTLA-4 protein binding to human B7-1/CD80

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Avi-tag Biotinylated Recombinant Human CTLA-4 Fc protein (AVI7268) was immobilized on a Biacore Sensor Chip CM5 via the Avi-tag biotin, and binding to Recombinant Human B7-1/CD80 Fc protein (Catalog # 10133-B1) was measured at a concentration range between 82 pM and 21 nM. The double-referenced sensorgram was fit to a 1:1 binding model to determine the binding kinetics and affinity, with an affinity constant of KD=0.2511 nM.

Reconstitution Calculator

Background: B7-1/CD80

B7-1/CD80 and B7-2/CD86, together with their receptors CD28 and CTLA-4, constitute one of the dominant co-stimulatory pathways that regulate T- and B-cell responses (1). 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 (2-6). Mature human B7-1 consists of a 208 amino acid (aa) extracellular domain (ECD) with two immunoglobulin-like domains, a 21 aa transmembrane domain, and a 25 aa cytoplasmic domain (7). Within the ECD, human B7-1 shares 50% aa sequence identity with mouse B7-1. Alternative splicing generates a 30 kDa soluble isoform that lacks the transmembrane segment and retains the ability to bind CD28 and CTLA-4 and an isoform that lacks the second Ig-like domain and the transmembrane segment (8). Both human and mouse B7-1 and B7-2 can bind to either human or mouse CD28 and CTLA-4 (1). 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 (2).

References

Ville, S. et al. (2015) Front. Immunol. 6:411.
Azuma, M. et al. (1993) Nature 366:76.
Freeman, G.J. et al. (1993) Science 262:909.
Chen, C. et al. (1994) J. Immunol. 152:4929.
Freeman, G.J. et al. (1993) J. Exp. Med. 178:2185
Lanier, L. et al. (1995) J. Immunol. 154:97.
Freeman, G.J. et al. (1989) J. Immunol. 143:2714.
Kakoulidou, M. et al. (2007) Scand. J. Immunol. 66:529.

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