Human PD-L1, Fc Tag (HPLC-verified) (PD1-H5258) is expressed from human 293 cells (HEK293)。 It contains AA Phe 19 - Arg 238 (Accession # NP_054862。1)。
Predicted N-terminus: Phe 19
This protein carries a human IgG1 Fc tag at the C-terminus。
The protein has a calculated MW of 51.4 kDa. The protein migrates as 60-75 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Less than 0.1 EU per μg by the LAL method.
>95% as determined by SDS-PAGE.
>90% as determined by SEC-HPLC.
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
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Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
No activity loss was observed after storage at:
- 4-8°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Human PD-L1, Fc Tag (HPLC-verified) on SDS-PAGE under reducing (R) condition。 The gel was stained overnight with Coomassie Blue。 The purity of the protein is greater than 95%。
The purity of Human PD-L1, Fc Tag (HPLC-verified) (Cat。 No。 PD1-H5258) was greater than 90% as determined by SEC-HPLC。
Immobilized Human PD-1, Strep Tag (Cat. No. PD1-H5284) at 2 μg/mL (100 μL/well) can bind Human PD-L1, Fc Tag (HPLC-verified) (Cat. No. PD1-H5258) with a linear range of 4-63 ng/mL (QC tested).
Immobilized Human PD-L1, Fc Tag (HPLC-verified) (Cat. No. PD1-H5258) at 5 μg/mL (100 μL/well) can bind Anti-PD-L1 MAb with a linear range of 0.2-2 ng/mL (Routinely tested).
Immobilized Human B7-1, Mouse IgG2a Fc Tag, low endotoxin (Cat。 No。 B71-H52A4) at 5 μg/mL (100 μL/well) can bind Human PD-L1, Fc Tag (HPLC-verified) (Cat。 No。 PD1-H5258) with a linear range of 0。01-0。313 μg/mL (Routinely tested)。
Serial dilutions of Anti-PD-L1 Neutralizing Antibody were added into Human PD-L1, Fc Tag (HPLC-verified) (Cat。 No。 PD1-H5258): Biotinylated Human PD-1, Fc,Avitag,His Tag (Cat。 No。 PD1-H82F2) binding reactions。 The half maximal inhibitory concentration (IC50) is 0。2966 μg/mL (Routinely tested)。
Loaded Human PD-1, His Tag (HPLC verified) (Cat。 No。 PD1-H5221) on HIS1K Biosensor, can bind Human PD-L1, Fc Tag (HPLC-verified) (Cat。 No。 PD1-H5258) with an affinity constant of 38。9 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested)。
活性（Bioactivity）-Cell based assay
Immobilized cell surface PD-1 (5x104 of cells per well) can bind Human PD-L1, Fc Tag (HPLC-verified) (Cat. No. PD1-H5258) with an EC50 of 0.029 μg/mL (Routinely tested).
Flow Cytometry assay shows that Human PD-L1, Fc Tag (HPLC-verified) (Cat。 No。 PD1-H5258) can bind to 293 cell overexpressing human PD-1。 The concentration of PD-L1 used is 10 μg/mL (Routinely tested)。
FACS analysis shows that the binding of Human PD-L1, Fc Tag (HPLC-verified) to 293 overexpressing PD-1 was inhibited by increasing concentration of neutralizing Anti-PD-L1 antibody. The concentration of PD-L1 used is 10 μg/mL. The IC50 is 12.92 μg/mL (Routinely tested).
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。