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Your Position: ホーム > Protein > B7-1 > B71-H82F2

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

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Based on the high binding affinity between biotin-streptavidin systems, our Streptavidin series products are recommended to use together with biotinylated proteins to get lower non-specific binding results in a shorter time.

  • 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
    Online(Val 35 - Asn 242) NP_005182.1

    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. As a result of glycosylation, the protein migrates as 65-95 kDa under reducing (R) condition, and 130-160 kDa under non-reducing (NR) condition (SDS-PAGE).

  • Biotinylation
    Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
  • 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. Normally trehalose is added as protectant before lyophilization.

    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
Biotinylated Human B7-1, Fc,Avitag (Cat. No. B71-H82F2) SDS-PAGE gel

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
Biotinylated Human B7-1, Fc,Avitag (Cat. No. ) 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
Biotinylated Human B7-1, Fc,AvitagBiotinylated Human B7-1, Fc,Avitag (Cat. No. B71-H82F2) ELISA bioactivity

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).

Biotinylated Human B7-1, Fc,AvitagBiotinylated Human B7-1, Fc,Avitag (Cat. No. B71-H82F2) ELISA bioactivity

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).

Biotinylated Human B7-1, Fc,AvitagBiotinylated Human B7-1, Fc,Avitag (Cat. No. B71-H82F2) ELISA bioactivity

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
FACS assay shows that recombinant Biotinylated 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).

FACS assay shows that recombinant Biotinylated 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
      
  • Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.

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