分子别名(Synonym)
PDCD1,PD1,CD279,SLEB2
表达区间及表达系统(Source)
Mouse PD-1, Fc Tag (PD1-M5259) is expressed from human 293 cells (HEK293). It contains AA Leu 25 - Gln 167 (Accession # NP_032824).
Predicted N-terminus: Leu 25
Request for sequence
蛋白结构(Molecular Characterization)
This protein carries a human IgG1 Fc tag at the C-terminus.
The protein has a calculated MW of 42.8 kDa. The protein migrates as 55-66 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
内毒素(Endotoxin)
Less than 1.0 EU per μg by the LAL method.
纯度(Purity)
>95% as determined by SDS-PAGE.
制剂(Formulation)
Lyophilized from 0.22 μm filtered solution in Tris with Glycine, Arginine and NaCl, pH7.5 with trehalose as protectant.
Contact us for customized product form or formulation.
重构方法(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.
存储(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.
电泳(SDS-PAGE)
Mouse PD-1, Fc Tag on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95%.
活性(Bioactivity)-ELISA
Immobilized Mouse PD-L1, Mouse IgG2a Fc Tag, low endotoxin (Cat. No. PD1-M52A2) at 2 μg/mL (100 μL/well) can bind Mouse PD-1, Fc Tag (Cat. No. PD1-M5259) with a linear range of 0.039-0.625 μg/mL (QC tested).
Protocol
活性(Bioactivity)-SPR
Captured Mouse PD-1, Fc Tag (Cat. No. PD1-M5259) on Protein A Chip can bind Mouse PD-L1, His Tag (Cat. No. PD1-M5220) with an affinity constant of 2.16 μM as determined in SPR assay (Biacore T200) (Routinely tested).
Protocol
Captured Mouse PD-L1, His Tag (Cat. No. PD1-M5220) on CM5 Chip via Anti-His antibody can bind Mouse PD-1, Fc Tag (Cat. No. PD1-M5259) with an affinity constant of 1.37 μM as determined in SPR assay (Biacore T200) (Routinely tested).
Protocol
活性(Bioactivity)-BLI
Loaded Mouse PD-1, Fc Tag (Cat. No. PD1-M5259) on Protein A Biosensor, can bind Mouse PD-L1, His Tag (Cat. No. PD1-M5220) with an affinity constant of 1.5 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
Protocol
Loaded Mouse PD-1, Fc Tag (Cat. No. PD1-M5259) on Protein A Biosensor, can bind Mouse PD-L2, His Tag (Cat. No. PD2-M52E3) with an affinity constant of 0.13 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
Protocol
背景(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.