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Your Position: 홈 > Protein > PD-1 > PD1-C82E6

Biotinylated Cynomolgus PD-1 / PDCD1 Protein, His,Avitag™

INTRODUCTION
QC & Assay Protocol
Citation & Background
  • Synonym
    PDCD1,PD1,CD279,SLEB2
  • Source
    Biotinylated Cynomolgus PD-1, His,Avitag(PD1-C82E6) is expressed from human 293 cells (HEK293). It contains AA Leu 25 - Gln 167 (Accession # B0LAJ3).
    Predicted N-terminus: Leu 25
    Request for sequence
  • Molecular Characterization
    PD-1 Structure

    This protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag (Avitag™).

    The protein has a calculated MW of 19.5 kDa. The protein migrates as 30-45 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.

  • Labeling
    Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
  • Protein Ratio
    Passed as determined by the HABA assay / binding ELISA.
  • Endotoxin
    Less than 1.0 EU per μg by the LAL method.
  • Purity

    >90% as determined by SDS-PAGE.

  • Formulation

    Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.

    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.

    This product is stable after storage at:

    1. -20°C to -70°C for 12 months in lyophilized state;
    2. -70°C for 3 months under sterile conditions after reconstitution.
SDS-PAGE
PD-1 SDS-PAGE

Biotinylated Cynomolgus PD-1, His,Avitag on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90%.

Bioactivity-ELISA
PD-1 ELISA

Immobilized Biotinylated Cynomolgus PD-1, His,Avitag (Cat. No. PD1-C82E6) at 1 μg/mL (100 μL/well) on streptavidin precoated (0.5 μg/well) plate, can bind Cynomolgus / Rhesus macaque PD-L1, Fc Tag (Cat. No. PD1-C5253) with a linear range of 1-5 ng/mL (QC tested).

Protocol
PD-1 ELISA

Immobilized Biotinylated Cynomolgus PD-1, His,Avitag (Cat. No. PD1-C82E6) at 1 μg/mL (100 μL/well) on Streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate, can bind Nivolumab with a linear range of 1-20 ng/mL (Routinely tested).

Protocol
PD-1 ELISA

Immobilized Human PD-L2, Fc Tag (HPLC verified) (Cat. No. PD2-H5251) at 5 μg/mL (100 μL/well) can bind Biotinylated Cynomolgus PD-1, His,Avitag (Cat. No. PD1-C82E6) with a linear range of 0.1-1.25 μg/mL (Routinely tested).

Protocol
Bioactivity-BLI
PD-1 BLI

Loaded Biotinylated Cynomolgus PD-1, His,Avitag (Cat. No. PD1-C82E6) on SA Biosensor, can bind Cynomolgus / Rhesus macaque PD-L1, His Tag (Cat. No. PD1-C52H4) with an affinity constant of 2.8 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
PD-1 BLI

Loaded Biotinylated Cynomolgus PD-1, His,Avitag (Cat. No. PD1-C82E6) on SA Biosensor, can bind Human PD-L2, His Tag (SPR verified) (Cat. No. PD2-H5220) with an affinity constant of 0.49 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
  • Background
    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.
  • Clinical and Translational Updates

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