Recombinant Human CD274 protein, mouse IgG1 Fc-tagged, low endotoxin (HPLC-verified) CD274-2146H

Recombinant Human CD274 protein, mouse IgG1 Fc-tagged, low endotoxin (HPLC-verified)

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Recombinant Human CD274 protein, mouse IgG1 Fc-tagged, low endotoxin (HPLC-verified)

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Cat.No. : CD274-2146H
Product Overview : Recombinant Human CD274 protein (Phe19-Arg238)(low endotoxin, HPLC-verified), fused to Mouse IgG1 Fc tag at C-terminus, was expressed in human 293 cells (HEK293).
Description : Programmed cell death 1 ligand 1 (PD-L1) is also known as cluster of differentiation (CD274) or B7 homolog 1 (B7-H1), is a member of the growing B7 family of immune molecules and is involved in the regulation of cellular and humoral immune responses. B7-H1 is a cell surface immunoglobulin superfamily with two Ig-like domains within the extracellular region and a short cytoplasmic domain. PD-L1 is highly expressed in the heart, skeletal muscle, placenta and lung and weakly expressed in the thymus, spleen, kidney and liver. PD-L1 is expressed on activated T-cells, B-cells, dendritic cells, keratinocytes and monocytes. PD-L1 is up-regulated on T- and B-cells, dendritic cells, keratinocytes and monocytes after LPS and IFNG activation and up-regulated in B-cells activated by surface Ig cross-linking. PD-L1 involve in the costimulatory signal, essential for T-cell proliferation and production of IL10 and IFNG, in an IL2-dependent and a PDCD1-independent manner.
Source : HEK293
Species : Human
Tag : Mouse IgG1 Fc
Form : Lyophilized from 0.22 um filtered solution in PBS, pH7.4, 10% trehalose.
Molecular Mass : The protein has a calculated MW of 51.6 kDa. As a result of glycosylation, the protein migrates as 65-85 kDa under reducing (R) condition, and 130-170 kDa under non-reducing (NR) condition (SDS-PAGE).
Protein length : 220
Endotoxin : Less than 0.1 EU per ug by the LAL method.
Purity : >92% as determined by SDS-PAGE.
Storage : For long term storage, the product should be stored at lyophilized state at -20 centigrade or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
-20 centigrade to -70 centigrade for 12 months in lyophilized state;
-70 centigrade for 3 months under sterile conditions after reconstitution.
Reconstitution : It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4ºC before opening to recover the entire contents.
SDS-PAGE of CD274-2146H :

Human PD-L1, Mouse IgG1 Fc Tag, low endotoxin (HPLC-verified) 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 92%.

SEC-HPLC of CD274-2146H :

The purity of Human PD-L1, Mouse IgG1 Fc Tag, low endotoxin (HPLC-verified) was more than 90% as determined by SEC-HPLC.

Bioactivity-ELISA of CD274-2146H :

Immobilized Human PD-1, Tag Free at 0.2 μg/mL (100 μL/well) can bind pre-mixed increasing concentrations of Nivolumab and 1 μg/mL (100 μL/well) Human PD-L1, Mouse IgG1 Fc Tag, low endotoxin (HPLC-verified) with a half maximal inhibitory concentration (IC50) of 0.30 μg/mL (QC tested).

Immobilized Human PD-1, Tag Free, Full Length at 0.2 μg/mL (100 μL/well) can bind Human PD-L1, Mouse IgG1 Fc Tag, low endotoxin (HPLC-verified) with a linear range of 5-156 ng/mL (Routinely tested).

Serial dilutions of nivolumab were added into Human PD-L1, Mouse IgG1 Fc Tag, low endotoxin (HPLC-verified): Biotinylated Human PD-1, Fc,Avitag binding reactions. The half maximal inhibitory concentration (IC50) is 0.3789 μg/mL (Routinely tested).

Bioactivity-FACS of CD274-2146H :

Flow Cytometry assay shows that Human PD-L1, Mouse IgG1 Fc Tag, low endotoxin (HPLC-verified) can bind to 293 cell overexpressing human PD-1. The concentration of PD-L1 used is 0.1 μg/mL (Routinely tested).

FACS analysis shows that the binding of Human PD-L1, Mouse IgG1 Fc Tag, low endotoxin (HPLC-verified) to 293 overexpressing PD-1 was inhibited by increasing concentration of neutralizing Anti-PD-1 antibody. The concentration of PD-L1 used is 1 μg/mL. The IC50 is 0.145 μg/mL (Routinely tested).

Gene Name : CD274
Official Symbol : CD274
Synonyms : CD274; CD274 molecule; CD274 antigen , PDCD1LG1, programmed cell death 1 ligand 1; programmed cell death 1 ligand 1; B7 homolog 1; B7 H; B7 H1; B7H1; PD L1; PDL1; CD274 antigen; PDCD1 ligand 1; programmed death ligand 1; B7-H; PD-L1; PDCD1L1; PDCD1LG1; MGC142294; MGC142296
Gene ID : 29126
mRNA Refseq : NM_014143
Protein Refseq : NP_054862
MIM : 605402
UniProt ID : Q9NZQ7

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