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Your Position: Home > Protein > B7-1 > B71-H82F2

Biotinylated Human B7-1 / CD80 Protein, Fc,Avitag™ (MALS verified)

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  • Synonym
    CD80,B7,B7-1,B7.1,BB1,CD28LG,CD28LG1,LAB7
  • Source
    Biotinylated Human B7-1, Fc,Avitag(B71-H82F2) is expressed from human 293 cells (HEK293). It contains AA Val 35 - Asn 242 (Accession # NP_005182.1).
    Predicted N-terminus: Val 35
  • Molecular Characterization
    B7-1 Structure

    This protein carries a human IgG1 Fc tag at the C-terminus, followed by an Avi tag (Avitag™).

    The protein has a calculated MW of 52.3 kDa. The protein migrates as 65-95 kDa under reducing (R) condition, and 130-160 kDa under non-reducing (NR) condition (SDS-PAGE) due to glycosylation.

  • Labeling
    Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
  • Protein Ratio
    Passed as determined by the HABA assay / binding ELISA.
  • Endotoxin
    Less than 1.0 EU per μg by the LAL method.
  • Purity

    >95% as determined by SDS-PAGE.

  • Formulation

    Lyophilized from 0.22 μm filtered solution in Tris with Glycine, Arginine and NaCl, pH7.5 with trehalose as protectant.

    Contact us for customized product form or formulation.

  • Reconstitution

    Please see Certificate of Analysis for specific instructions.

    For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

  • Storage

    For long term storage, the product should be stored at lyophilized state at -20°C or lower.

    Please avoid repeated freeze-thaw cycles.

    This product is stable after storage at:

    1. -20°C to -70°C for 12 months in lyophilized state;
    2. -70°C for 3 months under sterile conditions after reconstitution.
SDS-PAGE
B7-1 SDS-PAGE

Biotinylated Human B7-1, Fc,Avitag on SDS-PAGE under reducing (R) and non-reducing (NR) conditions. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95%.

SEC-MALS
B7-1 MALS images

The purity of Biotinylated Human B7-1, Fc,Avitag (Cat. No. B71-H82F2) is more than 85% and the molecular weight of this protein is around 130-165 kDa verified by SEC-MALS.

Bioactivity-ELISA
 B7-1 ELISA

Immobilized Human CTLA-4, Fc Tag (Cat. No. CT4-H5255) at 5 μg/mL (100 μL/well) can bind Biotinylated Human B7-1, Fc,Avitag (Cat. No. B71-H82F2) with a linear range of 0.6-9.8 ng/mL (QC tested).

 B7-1 ELISA

Immobilized Human PD-L1, Fc Tag (Cat. No. PD1-H5258) at 10 μg/mL (100 μL/well) can bind Biotinylated Human B7-1, Fc,Avitag (Cat. No. B71-H82F2) with a linear range of 0.156-5 μg/mL (Routinely tested).

 B7-1 ELISA

Serial dilutions of Ipilimumab were added into Human CTLA-4, Fc Tag (Cat. No. CT4-H5255): Biotinylated Human B7-1, Fc,Avitag (Cat. No. B71-H82F2) binding reactions. The half maximal inhibitory concentration (IC50) is 0.8260 μg/mL (Routinely tested).

Bioactivity-FACS
 B7-1 FACS

Flow Cytometry assay shows that Biotinylated Human B7-1, Fc,Avitag (Cat. No. B71-H82F2) can bind to 293 cell overexpressing human CTLA-4. The concentration of Human B7-1 is 0.3 μg/mL (Routinely tested).

 B7-1 FACS

FACS analysis shows that the binding of Biotinylated Human B7-1, Fc,Avitag (Cat. No. B71-H82F2) to 293 overexpressing CTLA-4 was inhibited by increasing concentration of neutralizing Anti-Human CTLA-4 antibody. The concentration of B7-1 used is 0.3 μg/mL. The IC50 is 3.025 μg/mL (Routinely tested).

  • Background
    B7-1 and B7-2, together with their receptors CD28 and CTLA­4, constitute one of the dominant co-stimulatory pathways that regulate T­ and B­cell responses. Although both CTLA­4 and CD28 can bind to the same ligands, CTLA­4 binds to B7­1 and B7­2 with a 20 ­ 100 fold higher affinity than CD28 and is involved in the down­regulation of the immune response.
    B-lymphocyte activation antigen B7-1 (referred to as B7) also known as cluster of Differentiation 80 (CD80), is a member of cell surface immunoglobulin superfamily and is expressed on activated B cells, activated T cells, macrophages and dendritic cells. It is the ligand for two different proteins on the T cell surface: CD28 (for autoregulation and intercellular association) and CTLA-4 (for attenuation of regulation and cellular disassociation). CD80 works in tandem with CD86 to prime T cells. CD80 plays a role in induction of innate immune responses by activating NF-κB-signaling pathway in macrophages. CD80 is thus regarded as promising therapeutic targets for autoimmune diseases and various carcinomas.
  • Clinical and Translational Updates

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