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Recombinant human B7-2 / CD86 Protein, Fc tag, 200µg  

Recombinant human B7-2 / CD86 Protein, Fc tag, 200µg

Recombinant Human B7-2 / CD86 (Leu 26 - Pro 247), expressed from human 293 cells (HEK293), Fc tag

Synonym
CD86, B7-2, B70, CD28LG2, LAB72, MGC34413

More details

CD6-H5257-200

Availability: within 7 days

408,00 €

Background
Cluster of Differentiation 86 (CD86), also known as B-lymphocyte activation antigen B7-2, is a type I membrane protein that is a member of the immunoglobulin superfamily,[1] and is constitutively expressed on interdigitating dendritic cells, Langerhans cells, peripheral blood dendritic cells, memory B cells, and germinal center B cells. Additionally, B7-2 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.[2] 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.[3]

Source
Recombinant Human B7-2, Fc Tag (CD6-H5257) is expressed from human 293 cells (HEK293). It contains AA Leu 26 - Pro 247 (Accession # AAH40261).
Predicted N-terminus: Leu 26

Molecular Characterization
This protein carries a human IgG1 Fc tag at the C-terminus.
The protein has a calculated MW of 51.5 kDa. As a result of glycosylation, the protein migrates as 66-86 kDa under reducing (R) condition, and 120-140 kDa under non-reducing (NR) condition (SDS-PAGE).

Endotoxin
Less than 1.0 EU per μg of the rhCD86-Fc by the LAL method.

Purity
>95% as determined by SDS-PAGE.

Formulation
Lyophilized from 0.22 μm filtered solution in 50 mM tris, 100 mM glycine, pH7.0. Normally Mannitol or Trehalose are added as protectants before lyophilization.

Reconstitution
See Certificate of Analysis for details of reconstitution instruction and specific concentration.

Storage
Avoid repeated freeze-thaw cycles. No activity loss was observed after storage at:
In lyophilized state for 1 year (4°C-8°C); After reconstitution under sterile conditions for 1 month (4°C-8°C) or 3 months (-20°C to -70°C).

Bioactivity
Immobilized Human B7-2, Fc Tag (Cat. No. CD6-H5257) at 5 μg/mL (100 μL/well) can bind Biotinylated Human CTLA-4, Fc Tag, Avi Tag (Cat. No. CT4-H82F3) with a linear range of 0.6-10 ng/mL (QC tested).
Flow Cytometry assay shows that recombinant Human B7-2, Fc Tag (Cat.No. CD6-H5257) can bind to CD28 expressed on Jurkat E6.1. The concentration of B7-2 used is 2 μg/ml(Routinely tested).
FACS analysis shows that the binding of Human B7-2, Fc Tag to CD28 expressed on Jurkat E6.1 was inhibited by increasing concentration of neutralizing  anti-CD28 antibody. The concentration of B7-2 used is 2 μg/ml. The IC50 is 0.022 μg/ml (Routinely tested).
FACS assay shows that Human B7-2, Fc Tag (Cat. No. CD6-H5257) can bind to 293T cells overexpressing human CTLA4 protein. The concentration of Human B7-2 is 3 μg/ml (Routinely tested).

References

(1) Chen C, et al., 1994, J. Immunol. 152 (10): 4929–36.
(2) Yadav, D. et al., 2007, J. Immunol. 178: 6236-6241.
(3) Steidl C, et al., 2010, N. Engl. J. Med. 362 (10): 875–85.