分子别名（Synonym）
PDCD1,PD1,CD279,SLEB2
表达区间及表达系统（Source）
Human PD-1, Fc Tag, low endotoxin (PD1-H5257) is expressed from human 293 cells (HEK293). It contains AA Leu 25 - Gln 167 (Accession # NP_005009.2).
N-terminal Sequence Analysis: Leu 25
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 42.1 kDa. The protein migrates as 56-66 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
内毒素（Endotoxin）
Less than 0.1 EU per μg by the LAL method.
纯度（Purity）
>95% as determined by SDS-PAGE.
>90% as determined by SEC-MALS.
制剂（Formulation）
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.
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）
Human PD-1, Fc Tag, low endotoxin on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
SEC-MALS
The purity of Human PD-1, Fc Tag, low endotoxin(Cat. No. PD1-H5257) was more than 90% and the molecular weight of this protein is around 130-150 kDa verified by SEC-MALS.
Report
活性（Bioactivity）-ELISA
Immobilized Biotinylated Human PD-L1, Avitag,His Tag (recommended for biopanning) (Cat. No. PD1-H82E5) at 1 μg/mL (100 μL/well) on streptavidin precoated (0.2 μg/well) plate, can bind Human PD-1, Fc Tag, low endotoxin (Cat. No. PD1-H5257) with a linear range of 0.1-3 ng/mL (QC tested).
Protocol
Report
Immobilized Human PD-1, Fc Tag, low endotoxin (Cat. No. PD1-H5257) at 2 μg/mL (100 μL/well) can bind Biotinylated Human PD-L2, Fc,Avitag (Cat. No. PD2-H82F6) with a linear range of 0.5-16 ng/mL (Routinely tested).
Protocol
Immobilized Biotinylated Human PD-L2, Avitag,His Tag (recommended for biopanning) (Cat. No. PD2-H82E8) at 1 μg/mL (100 μL/well) on streptavidin precoated (0.2 μg/well) plate, can bind Human PD-1, Fc Tag, low endotoxin (Cat. No. PD1-H5257) with a linear range of 0.1-1 ng/mL (Routinely tested).
Protocol
Serial dilutions of nivolumab were added into Human PD-1, Fc Tag, low endotoxin (Cat. No. PD1-H5257): Biotinylated Human PD-L1, Fc,Avitag,His Tag (Cat. No. PD1-H82F3) binding reactions. The half maximal inhibitory concentration (IC50) is 0.5381 μg/mL (Routinely tested).
Protocol
活性（Bioactivity）-SPR
Human PD-1, Fc Tag, low endotoxin (HPLC-verified) (Cat. No. PD1-H5257) captured on CM5 chip via anti-human IgG Fc antibody, can bind Human PD-L1, His Tag (HPLC-verified) (Cat. No. PD1-H5229) with an affinity constant of 3.6 μM as determined in a SPR assay (Biacore T200) (Routinely tested).
Protocol
活性（Bioactivity）-BLI
Loaded Human PD-1, Fc Tag, low endotoxin (Cat. No. PD1-H5257) on ProteinA Biosensor, can bind Human PD-L1, His Tag (HPLC & BLI verified) (Cat. No. PD1-H5229) with an affinity constant of 5.3 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
Protocol
Loaded Human PD-1, Fc Tag, low endotoxin (Cat. No. PD1-H5257) on Protein A Biosensor, can bind Human PD-L2, His Tag (SPR verified) (Cat. No. PD2-H5220) with an affinity constant of 0.45 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
Protocol
活性（Bioactivity）-FACS
Flow Cytometry assay shows that Human PD-1, Fc Tag, low endotoxin (Cat. No. PD1-H5257) can bind to 293 cell overexpressing human PD-L1. The concentration of PD-1 used is 1 μg/mL (Routinely tested).
Protocol
文献引用（Citations）
Heterocyclic compounds as immunomodulators
Authors: L Wu, J Li, W Yao
Journal: US10669271B2 2020
Application: Binding Assay
Authors: Wang Y, Ni H, Zhou S, et al.
Journal: Cancer Immunol Immunother 2020
Application: Flow cytometry
Heterocyclic compounds as immunomodulators
Authors: L Wu, F Zhang, S Mei, et al.
Journal: US10669271B2 2020
Application: Binding Assay
Authors: Zhao Y, et al
Journal: Immunity 2019
Application: Cell Culture
Comprehensive in vitro characterization of PD-L1 small molecule inhibitors
Authors: Ganesan A, et al
Journal: Sci Rep 2019
Application: ELISA
Artificial T cell mimetics to combat melanoma tumor growth
Authors: Mukundan S, et al.
Journal: Am J Adv Drug Deliv 2018
Application: Flow Cytometry
Immunomodulator compounds and methods of use
Authors: P Liu, et al.
Journal: US20180177870A1 2018
Application: Alphascreen & HTRF assay
PD-L1 Specific Monoclonal Antibodies for Disease Treatment and Diagnosis
Authors: Wang Aijun, et al.
Journal: US20170355770A1 2017
Application: ELISA
Heterocyclic compounds as immunomodulators
Authors: Zhenwu Li, et al.
Journal: US20170320875A1 2017
Application: HTRF Binding Assay
Hererocyclic compounds as immunomodulators
Authors: J Li, et al.
Journal: US20170145025A1 2017
Application: HTRF Binding Assay
Hererocyclic compounds as immunomodulators
Authors: L Wu, et al.
Journal: US20170107216A1 2017
Application: HTRF Binding Assay
A novel PD-L1-targeting antagonistic DNA aptamer with antitumor effects.
Authors: Lai WY, et al.
Journal: Mol Ther Nucleic Acids 2016
Application: ELISA-based competition assay
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.
