Biotinylated Human B7-2, Fc,Avitag (CD6-H82F5) is expressed from human 293 cells (HEK293). It contains AA Leu 26 - Pro 247 (Accession # AAH40261).
Predicted N-terminus: Leu 26
This protein carries a human IgG1 Fc tag at the C-terminus, followed by a Avi tag (Avitag™).
The protein has a calculated MW of 53.8 kDa. As a result of glycosylation, the protein migrates as 66-100 kDa under reducing (R) condition, and 120-140 kDa under non-reducing (NR) condition (SDS-PAGE).
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
The biotin to protein ratio is 0.5-1 as determined by the HABA assay.
Less than 1.0 EU per μg by the LAL method.
>95% as determined by SDS-PAGE.
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.
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
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:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human B7-2, Fc,Avitag on SDS-PAGE under reducing (R) and non-reducing (NR) conditions. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
Immobilized Human CTLA-4, Fc Tag (Cat. No. CT4-H5255) at 5 μg/mL (100 μL/well) can bind Biotinylated Human B7-2, Fc,Avitag (Cat. No. CD6-H82F5) with a linear range of 19-156 ng/mL (QC tested).
Serial dilutions of Ipilimumab were added into Human CTLA-4, Fc Tag (Cat. No. CT4-H5255): Biotinylated Human B7-2, Fc,Avitag (Cat. No. CD6-H82F5) binding reactions. The half maximal inhibitory concentration (IC50) is 0.1701 μg/mL (Routinely tested).
Flow Cytometry assay shows that Biotinylated Human B7-2, Fc,Avitag (Cat. No. CD6-H82F5) can bind to CD28 expressed on Jurkat E6.1 cells. The concentration of B7-2 used is 1.5 μg/mL (Routinely tested).
FACS analysis shows that the binding of Biotinylated Human B7-2, Fc,Avitag to CD28 expressed on Jurkat E6.1 was inhibited by increasing concentration of neutralizing Anti-CD28 antibody. The concentration of B7-2 used is 1.5 μg/mL. The IC50 is 0.031 μg/mL (Routinely tested).
Flow Cytometry assay shows that Biotinylated Human B7-2, Fc,Avitag (Cat. No. CD6-H82F5) can bind to 293 cell overexpressing human CTLA-4. The concentration of Human B7-2 is 2 μg/mL (Routinely tested).
FACS analysis shows that the binding of Biotinylated Human B7-2, Fc,Avitag (Cat. No. CD6-H82F5) to 293 overexpressing CTLA-4 was inhibited by increasing concentration of neutralizing Anti-human CTLA-4 antibody. The concentration of Human B7-2 is 2 μg /mL. The IC50 is 1.014 μg/mL (Routinely tested).
Cluster of Differentiation 86 (CD86) is also known as B-lymphocyte activation antigen B7-2, is a type I membrane protein that is a member of the immunoglobulin superfamily, and is constitutively expressed on interdigitating dendritic cells, Langerhans cells, peripheral blood dendritic cells, memory B cells, and germinal center B cells. Additionally, B72 is expressed at low levels on monocytes and can be upregulated through interferon γ. CD86 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). CD86 works in tandem with CD80 to prime T cells. Recent study has revealed that B7-2 promotes the generation of a mature APC repertoire and promotes APC function and survival. Furthermore, the B7 proteins are also involved in innate immune responses by activating NF-κB-signaling pathway in macrophages. CD86 thus is regarded as a promising candidate for immune therapy. CD86+ macrophages in Hodgkin lymphoma patients are an independent marker for potential nonresponse to firstline-therapy.