MABSol® Biotinylated Human PD-1, Fc,Avitag,His Tag (PD1-H82F2) is expressed from human HEK293 cells. It contains AA Leu 25 - Gln 167 (Accession # Q15116-1).
Predicted N-terminus: Leu 25
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 44.7 kDa. The protein migrates as 50-66 kDa on a SDS-PAGE gel under reducing (R) condition due to glycosylation and 100-130 kDa under non-reducing (NR) condition.
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 PBS, pH7.4. 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 PD-1, Fc,Avitag,His Tag 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 PD-L1, Fc Tag (HPLC-verified) (Cat. No. PD1-H5258) at 5 μg/mL (100 μL/well) can bind Biotinylated Human PD-1, Fc,Avitag,His Tag (Cat. No. PD1-H82F2) with a linear range of 16-125 ng/mL (QC tested).
Immobilized Nivolumab at 2 μg/mL (100 μL/well) can bind Biotinylated Human PD-1, Fc,Avitag,His Tag (Cat. No. PD1-H82F2) with a linear range of 0.4-6 ng/mL (Routinely tested).
Serial dilutions of Anti-PD-L1 Neutralizing Antibody were added into Human PD-L1, Fc Tag (HPLC-verified) (Cat. No. PD1-H5258): Biotinylated Human PD-1, Fc,Avitag,His Tag (Cat. No. PD1-H82F2) binding reactions. The half maximal inhibitory concentration (IC50) is 0.2966 μg/mL (Routinely tested).
Loaded Biotinylated Human PD-1, Fc,Avitag,His Tag (Cat. No. PD1-H82F2) on SA Biosensor, can bind Human PD-L1, His Tag (HPLC verified) (Cat. No. PD1-H5229) with an affinity constant of 2.2 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
Loaded Biotinylated Human PD-1, Fc,Avitag,His Tag (Cat. No. PD1-H82F2) on SA Biosensor, can bind Human PD-L2, His Tag (SPR verified) (Cat. No. PD2-H5220) with an affinity constant of 0.36 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
Flow Cytometry assay shows that recombinant Biotinylated Human PD-1, Fc,Avitag,His Tag (Cat No. PD1-H82F2) can bind to 293 cell overexpressing human PD-L1. The concentration of PD-1 used is 1 μg/mL (Routinely tested).
FACS analysis shows that the binding of Biotinylated Human PD-1, Fc,Avitag,His Tag to 293 overexpressing PD-L1 was inhibited by increasing concentration of neutralizing anti-PD-L1 antibody. The concentration of PD-1 used is 1 μg/mL. The IC50 is 0.362 μg/mL (Routinely tested).
Programmed cell death protein 1 (PD-1) is also known as CD279 and PDCD1, is a type I membrane protein and is a member of the extended CD28/CTLA-4 family of T cell regulators. PDCD1 is expressed on the surface of activated T cells, B cells, macrophages, myeloid cells and a subset of thymocytes. PD-1 has two ligands, PD-L1 and PD-L2, which are members of the B7 family. PD-L1 is expressed on almost all murine tumor cell lines, including PA1 myeloma, P815 mastocytoma, and B16 melanoma upon treatment with IFN-γ. PD-L2 expression is more restricted and is expressed mainly by DCs and a few tumor lines. PD1 inhibits the T-cell proliferation and production of related cytokines including IL-1, IL-4, IL-10 and IFN-γ by suppressing the activation and transduction of PI3K/AKT pathway. In addition, coligation of PD1 inhibits BCR-mediating signal by dephosphorylating key signal transducer. In vitro, treatment of anti-CD3 stimulated T cells with PD-L1-Ig results in reduced T cell proliferation and IFN-γ secretion. Monoclonal antibodies targeting PD-1 that boost the immune system are being developed for the treatment of cancer.