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INTRODUCTION
QC & Assay Protocol
Citation & Background
Synonym
PDCD1,PD1,CD279,SLEB2
Source
Mouse PD-1, His Tag(PD1-M5228) is expressed from human 293 cells (HEK293). It contains AA Leu 25 - Gln 167 (Accession # NP_032824).
Predicted N-terminus: Leu 25
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Molecular Characterization
This protein carries a polyhistidine tag at the C-terminus.
The protein has a calculated MW of 17.0 kDa. The protein migrates as 27-43 kDa when calibrated against Star Ribbon Pre-stained Protein Marker under reducing (R) condition (SDS-PAGE) due to different glycosylation.
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.
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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:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
SDS-PAGE
Mouse PD-1, His Tag on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90% (With Star Ribbon Pre-stained Protein Marker).
Bioactivity-BLI
Loaded Mouse PD-L1, Fc Tag (Cat. No. PD1-M5251) on Protein A Biosensor, can bind Mouse PD-1, His Tag (Cat. No. PD1-M5228) with an affinity constant of 6.9 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
Protocol
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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.
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Clinical and Translational Updates
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Datasheet & Documentation
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