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Your Position: Home > Immune Checkpoint Proteins > PD-1 > PD1-HP2F2

PE-Labeled Human PD-1 / PDCD1 Protein, Fc,His Tag (recommended for neutralizing assay)

INTRODUCTION
QC & Assay Protocol
Citation & Background
  • Synonym
    PDCD1,PD1,CD279,SLEB2
  • Source
    PE-Labeled Human PD-1, Fc,His Tag (Cat. No. PD1-HP2F2) is produced via site-specific conjugation of PE to Human PD-1, Fc,His Tag under optimal conditions with a proprietary technology. Human PD-1, Fc,His Tag is expressed from human 293 cells (HEK293). It contains AA Leu 25 - Gln 167 (Accession # Q15116-1).
    Predicted N-terminus: Leu 25
    Request for sequence
  • Molecular Characterization
    Online(Leu 25 - Gln 167) Q15116-1

    This protein carries a human IgG1 Fc tag at the C-terminus, followed by a polyhistidine tag.

    The protein has a calculated MW of 44.7 kDa.

  • Application
    Flow Cytometry (Neutralizing assay)
  • Conjugate

    PE

    Excitation source: 488 nm spectral line, argon-ion laser

    Excitation Wavelength: 488 nm

    Emission Wavelength: 575 nm

  • Formulation

    Lyophilized from 0.22 μm filtered solution in PBS, 0.5% BSA, 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 protect from light and avoid repeated freeze-thaw cycles.

    This product is stable after storage at:

    1. -20°C to -70°C for 12 months in lyophilized state;
    2. -70°C for 3 months under sterile conditions after reconstitution.
Bioactivity-FACS
FACS assay shows that recombinant Biotinylated FACS

Flow Cytometry assay shows that PE-Labeled Human PD-1, Fc,His Tag (Cat. No. PD1-HP2F2) can bind to 293T cells overexpressing human PD-L1. 1 μL stock solution per million cells (QC tested).

Protocol
FACS assay shows that recombinant Biotinylated FACS

FACS analysis shows that the binding of PE-Labeled Human PD-1, Fc,His Tag (Cat. No. PD1-HP2F2) to 293T overexpressing PD-L1 was inhibited by increasing concentration of neutralizing anti-human PD-L1 antibody. The IC50 is 0.29 μg/mL (QC tested).

Protocol
  • 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.
  • References
      

  • Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.

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