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Biotinylated Human B7-1 / CD80 Protein, Fc Tag, Avi Tag (Avitag™)  pdf  pdf  pdf


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B71-H82F2-25ug
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Synonym

CD80,B7,B7-1,B7.1,BB1,CD28LG,CD28LG1,LAB7

Source

Biotinylated Human B7-1, Fc Tag (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(Val 35 - Asn 242)NP_005182.1
Fc(Pro 100 - Lys 330)P01857
Avi

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

The biotin to protein ratio is 0.5-1 as determined by the HABA assay.

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 50 mM tris, 100 mM glycine, 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.

No activity loss is observed after storage at:

  1. 4-8°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 Tag (Catalog # B71-H82F2) SDS-PAGE gel

Biotinylated Human B7-1, Fc Tag on SDS-PAGE under reducing (R) and no-reducing (NR) conditions. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.

Bioactivity

Biotinylated Human B7-1, Fc Tag (Catalog # B71-H82F2) ELISA bioactivity

Immobilized Human CTLA-4, Fc Tag (Catalog # CT4-H5255) at 5 μg/mL (100 μL/well) can bind Biotinylated Human B7-1, Fc Tag (Catalog # B71-H82F2) with a linear range of 0.6-9.8 ng/mL.

Request for protocol

 

Background

B7-1 and B7-2, together with their receptors CD28 and CTLA¬4, constitute one of the dominant co-stimulatory pathways that regulate T¬ and B¬cell responses. Although both CTLA¬4 and CD28 can bind to the same ligands, CTLA¬4 binds to B7¬1 and B7¬2 with a 20 ¬ 100 fold higher affinity than CD28 and is involved in the down¬regulation 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.

References

 

Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.


Limited Use License

The Biotin AviTag technology is covered by U.S. Pat. No: 5,874,239 and includes any and all materials, methods, kits and related derivatives claimed by this patent. The purchase of the Acrobiosystems’s Avitag™ proteins confers to the purchaser the limited right to use the Avitag™ technology for non-commercial, or research use, or for purposes of evaluating the Avitag™ technology.

Commercial use of the Avitag™ technology to manufacture a commercial product, or use of the Avitag™ technology to facilitate or advance research which will be applied to the development of a commercial product requires a license from Avidity, LLC. Examples of Commercial use include, but are not limited to biosensors, diagnostics, therapeutic applications, proximity assays, and drug screening assays.