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Your Position: Home > Immune Checkpoint Proteins > PD-L1 > PD1-H5258

Human PD-L1 / B7-H1 Protein, Fc Tag (MALS verified)

INTRODUCTION
QC & Assay Protocol
Citation & Background
  • Synonym
    PD-L1,CD274,B7-H1,PDCD1L1,PDCD1LG1
  • Source
    Human PD-L1, Fc Tag (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
    Request for sequence
  • Molecular Characterization
    Online(Phe 19 - Arg 238) NP_054862.1

    This protein carries a human IgG1 Fc tag at the C-terminus.

    The protein has a calculated MW of 51.3 kDa. The protein migrates as 60-75 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.

  • Endotoxin
    Less than 0.1 EU per μg by the LAL method.
  • Purity

    >95% as determined by SDS-PAGE.

    >90% as determined by SEC-MALS.

  • 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.

  • 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:

    1. -20°C to -70°C for 24 months in lyophilized state;
    2. -70°C for 12 months under sterile conditions after reconstitution.
SDS-PAGE
Human PD-L1, Fc Tag (Cat. No. PD1-H5258) SDS-PAGE gel

Human PD-L1, Fc Tag 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
Human PD-L1, Fc Tag (Cat. No. ) MALS images

The purity of Human PD-L1, Fc Tag (Cat. No. PD1-H5258) was more than 90% and the molecular weight of this protein is around 115-150 kDa verified by SEC-MALS.

Report
Bioactivity-ELISA
Human PD-L1, Fc TagHuman PD-L1, Fc Tag (Cat. No. PD1-H5258) ELISA bioactivity

Immobilized Human PD-1, Strep Tag (Cat. No. PD1-H5284) at 2 μg/mL (100 μL/well) can bind Human PD-L1, Fc Tag (Cat. No. PD1-H5258) with a linear range of 4-63 ng/mL (QC tested).

Protocol
Human PD-L1, Fc TagHuman PD-L1, Fc Tag (Cat. No. PD1-H5258) ELISA bioactivity
Report
Human PD-L1, Fc TagHuman PD-L1, Fc Tag (Cat. No. PD1-H5258) ELISA bioactivity

Immobilized Human PD-L1, Fc Tag (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).

Protocol
Human PD-L1, Fc TagHuman PD-L1, Fc Tag (Cat. No. PD1-H5258) ELISA bioactivity

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 (Cat. No. PD1-H5258) with a linear range of 0.01-0.313 μg/mL (Routinely tested).

Protocol
Human PD-L1, Fc TagHuman PD-L1, Fc Tag (Cat. No. PD1-H5258) ELISA bioactivity

Serial dilutions of Anti-PD-L1 Neutralizing Antibody were added into Human PD-L1, Fc Tag (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).

Protocol
Bioactivity-BLI
Human_FcRn_Heterodimer_Protein_Bli

Loaded Human PD-1, His Tag (Cat. No. PD1-H5221) on HIS1K Biosensor, can bind Human PD-L1, Fc Tag (Cat. No. PD1-H5258) with an affinity constant of 38.9 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
Human_FcRn_Heterodimer_Protein_Bli

Loaded Human PD-L1, Fc Tag (Cat. No. PD1-H5258) on Protein A Biosensor, can bind Human B7-1, His Tag (Cat. No. B71-H5228) with an affinity constant of 18 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
Human_FcRn_Heterodimer_Protein_Bli

Loaded Human PD-L1, Fc Tag (Cat. No. PD1-H5258) on Protein A Biosensor, can bind Human Human PD-1, His Tag, low endotoxin (Cat. No. PD1-H522a) with an affinity constant of 4.8 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
Human_FcRn_Heterodimer_Protein_Bli

Loaded Human PD-L1, Fc Tag (Cat. No. PD1-H5258) on Protein A Biosensor, can bind Human Human PD-1, Strep Tag (Cat. No. PD1-H5284) with an affinity constant of 2.5 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
Bioactivity-FACS
FACS assay shows that recombinant Biotinylated FACS

Flow Cytometry assay shows that Human PD-L1, Fc Tag (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).

Protocol
FACS assay shows that recombinant Biotinylated FACS

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).

Protocol
Bioactivity-Cell based assay
Human FcRn Heterodimer Protein Cell_Base

Immobilized cell surface PD-1 (5x104 of cells per well) can bind Human PD-L1, Fc Tag (Cat. No. PD1-H5258) with an EC50 of 0.029 μg/mL (Routinely tested).

Protocol

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