Human CD80 Protein, ECD (extracellular domain), Fc-fusion, Biotinylated, Recombinant

Product ID: CD80-Fc-Biot (SKU#: FCL0723B)

Price:

$528.00

Description

CD80, also known as the B-cell activation antigen B7-1, is a single-pass type I transmemebrane glycoprotein that belongs to the B7 family of the Ig superfamily. Like other B7 family members, CD80 contains 1 Ig-like C2-type domain and 1 Ig-like V-type domain in the extracellular region. Among the B7 family members, they share about 20-25% amino acid identity. CD80 is expressed on the surface of antigen-presenting cells including activated B cells, macrophages and dendritic cells. CD80 provides a co-stimulatory signal necessary for T cell activation and survival. CD80 is the ligand for two different receptors on the T cell surface: CD28 and CTLA-4 (also known as CD152). CD80 works in concert with CD86 (also known as B7-2) to prime T cells. CD80 and CD86, together with their receptors CD28 and CTLA4, constitute one of the dominant co-stimulatory pathways that regulate T and B cell responses, tolerance and the generation of CTL. The binding of CD80:CD86 to its receptor CD28 induces T-cell proliferation and cytokine production. CTLA4 binds to CD80 and CD86 with a 20-100 fold higher affinity than CD28 and is involved in the downregulation of the immune response. CD80 also binds to B7 family ligand PD-L1 to regulate immune cell activation. CD80 may act as a receptor for adenovirus subgroup B and play a role in lupus neuropathy. CD80 is regarded as a promising therapeutic target for autoimmune diseases and cancer. Human CD80 and CD86 share 26% amino acid identity, while human and mouse CD80 share 44% amino acid identity. It has been reported that both human and mouse CD80 and CD86 can bind to either human or mouse CD28 and CTLA4.

Gene Symbol: CD80; B7-1; B7.1; BB1; BB-1; B7; CD28LG; CD28LG1; LAB7

NCBI Gene ID: 941

Uniprot Entry: P33681

Construct Details: The recombinant human CD80-Fc fusion protein is expressed as a 437amino acid protein consisting of Val35 - Asn242 region of CD80 (UniProt accession #P33681) and a C-terminal Fc fusion from human IgG1, which exists as a dimer under non-reducing condition.

Source: Human cells stably expressing CD80-Fc and growing in chemical-defined media with no animal components or antibiotics

Deduced Amino Acid Sequence:

VIHVTKEVKEVATLSCGHNVSVEELAQTRIYWQKEKKMVLTMMSGDMNIWPEYKNRTIFDITNNLSIVILALR

PSDEGTYECVVLKYEKDAFKREHLAEVTLSVKADFPTPSISDFEIPTSNIRRIICSTSGGFPEPHLSWLENGE

ELNAINTTVSQDPETELYAVSSKLDFNMTTNHSFMCLIKYGHLRVNQTFNWNTTKQEHFPDNGSTGTHTCPPC

PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV

VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS

DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

M.W.: Calculated molecular mass 49.5 kDa; estimated by SDS-PAGE under reducing condition 70-75 kDa probably due to glycosylation

Calculated PI: 6.00

Calculated Extinction Coefficients (M-1 cm-1, at 280nm): 66975

Purity: >95% judged by SDS-PAGE under reducing condition (see the gel image above)

Formulation: Supplied at 0.5 mg/ml in sterile PBSpH7.4 (carrier free & preservative free). The purified recombinant protein was labeled with Biotin (3-5 Biotin per molecule) using the standard procedure.

Endotoxin Level: <0.1 EU per 1 μg of purified recombinant protein determined by the LAL method

Biological Activity: Binds to human CD28, CTLA4 and PD-L1 by ELISA using immobilized CD80 protein. Binds to human and mouse PD-L1 (SKU#FCL0781, FCL1846) in a functional ELISA (see Technical Data). Induces IL-2 secretion by Jurkat human acute T cell leukemia cells with an ED₅₀ of 0.05 - 0.2 µg/mL in the presence of PHA.

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable. Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose. Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules. Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction. Your use of this product constitutes acceptance of the terms of this limited use agreement. Please refer to our “terms & conditions” for details. If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.

Technical Data 1: Interaction with Human PD-L1 by ELISA

Recombinant human PD-L1-Fc (SKU#FCL0781) was immobilized at 100 ng/well on a 96-well NuncMaxisorp plate for a functional ELISA with serially diluted, biotinylated human PD-1-Fc (PD-1-Fc-Biot, SKU#FCL0763B), biotinylated human CD80-Fc (CD80-Fc-Biot, SKU#FCL0723B), and biotinylated Fc alone (control Fc-Biot, SKU#FCL00029B). Fc protein alone was used as a negative control to demonstrate the specificity of PD-1:PD-L1 and CD80:PD-L1 interactions under the experimental conditions. The binding curves were fit and apparent KDwas calculated for PD-1: PD-L1 binding to be 365 ± 38 ng/ml with a linear binding range between 100 and 1500 ng/ml, and apparent KD for CD80:PD-L1 binding to be 831 ± 101 ng/ml with a linear binding range between 250 and 4000 ng/ml.

Technical Data 2: Cross-species Binding to Mouse PD-L1 by ELISA

Recombinant mouse PD-L1-Fc (SKU#FCL1846) was immobilized at 100 ng/well on a 96-well NuncMaxisorp plate for a functional ELISA with serially diluted, biotinylated human PD-1-Fc (PD-1-Fc-Biot, SKU#FCL0763B), biotinylated human CD80-Fc (CD80-Fc-Biot, SKU#FCL0723B), and biotinylated Fc alone (control Fc-Biot, SKU#FCL00029B). Fc protein alone was used as a negative control to demonstrate the specificity of PD-1:mPD-L1 and CD80:mPD-L1 interactions under the experimental conditions. Mouse PD-L1 shows cross-species reactivity to human PD-1 and CD80. The binding curve was fit and apparent K_D was calculated for PD-1: mPD-L1 binding to be 557 ± 63 ng/ml with a linear binding range between 150 and 2000 ng/ml, and apparent K_D for CD80:mPD-L1 binding to be 9486 ± 5500 ng/ml with a linear binding range between 1500 and 75000 ng/ml.

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

The product is shipped at 4°C. Upon receipt, centrifuge the product briefly before opening the vial. It is recommended to store small aliquots at the temperature below –20°C for long-term storage and the product is stable for 3 months. The undiluted protein can be stored at 4°C for no more than 2 weeks.

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

1. J. Exp. Med. 174:625 (1991).

2. Nature 366:76 (1993).

3. Science 262:909 (1993).

4. J. Immunol. 152:4929 (1994).

5. Annu. Rev. Immunol. 23:515 (2005).

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Product datasheet (pdf) can be downloaded here: FCL0723B-PDS.pdf