Biotinylated Human PD-L1, Fc,Avitag,His Tag (PD1-H82F3) is expressed from human 293 cells (HEK293). It contains AA Phe 19 - Arg 238 (Accession # NP_054862.1).
Predicted N-terminus: Phe 19
This protein carries a human IgG1 Fc fragment at the C-terminus, followed by a polyhistidine tag. The Avi tag (Avitag™) is inserted in-between the Fc and his tags. The protein has a calculated MW of 53.2 kDa. The protein migrates as 66-75 kDa on a SDS-PAGE gel under reducing (R) condition due to glycosylation.
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 50 mM Tris, 100 mM Glycine, 25 mM Arginine, 10 mM NaCl, pH7.5. Normally trehalose is added as protectant before lyophilization.
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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.
No activity loss was observed after storage at:
- 4-8°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human PD-L1, Fc,Avitag,His Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
Immobilized Human PD-1, Fc Tag (Cat. No. PD1-H5257) at 2 μg/mL (100μL/well) can bind Biotinylated Human PD-L1, Fc,Avitag,His Tag (Cat. No. PD1-H82F3) with a linear range of 0.01-0.25 μg/mL (QC tested).
Immobilized Human B7-1, mouse IgG2a Fc Tag, low endotoxin (Cat. No. B71-H52A4) at 5 μg/mL (100 μL/well) can bind Biotinylated Human PD-L1, Fc,Avitag,His Tag (Cat. No. PD1-H82F3) with a linear range of 0.02-0.625 μg/mL (Routinely tested).
活性（Bioactivity）-Cell based assay
Immobilized cell surface PD-1 (5x104 of cells per well) can bind Biotinylated Human PD-L1, Fc,Avitag,His Tag (Cat. No. PD1-H82F3) with an EC50 of 0.082 μg/mL (Routinely tested).
Flow Cytometry assay shows that Biotinylated Human PD-L1, Fc,Avitag,His Tag (Cat.No. PD1-H82F3) can bind to 293 cell overexpressing human PD-1. The concentration of PD-L1 used is 1 μg/mL (Routinely tested).
FACS analysis shows that the binding of Biotinylated Human PD-L1, Fc,Avitag,His Tag (Cat. No. PD1-H82F3) to 293 overexpressing PD-1 was inhibited by increasing concentration of neutralizing anti-PD-1 antibody. The concentration of PD-L1 used is 2 μg/mL. The IC50 is 0.64 μg/mL (Routinely tested).
Programmed cell death 1 ligand 1 (PD-L1) is also known as cluster of differentiation (CD274) or B7 homolog 1 (B7-H1), is a member of the growing B7 family of immune molecules and is involved in the regulation of cellular and humoral immune responses. B7-H1 is a cell surface immunoglobulin superfamily with two Ig-like domains within the extracellular region and a short cytoplasmic domain. PD-L1 is highly expressed in the heart, skeletal muscle, placenta and lung and weakly expressed in the thymus, spleen, kidney and liver. PD-L1 is expressed on activated T-cells, B-cells, dendritic cells, keratinocytes and monocytes. PD-L1 is up-regulated on T- and B-cells, dendritic cells, keratinocytes and monocytes after LPS and IFNG activation and up-regulated in B-cells activated by surface Ig cross-linking. PD-L1 involve in the costimulatory signal, essential for T-cell proliferation and production of IL10 and IFNG, in an IL2-dependent and a PDCD1-independent manner.