CD274 (B7-H1, PD-L1, Programmed Death Ligand) is a member of the B7 family and is expressed on a variety of tissues including lymphoid cells. It plays an important role in regulation of T cell activation, and is involved in progression of cancer, arthritis and HIV infection. CD274 binding to its receptor CD279 (PD-1) on activated T cells can decrease proliferation. Conversely, ligation of CD279 on primed T cells can stimulate IL-10 production. High levels of CD274 present in Renal cell carcinoma is associated with poor prognosis. Tumor expressed CD274 can increase apoptosis of tumor specific T cells resulting in better tumor cell survival. Gamma Interferon and PHA can up regulate CD274 expression on T cells.
Applications:
Suitable for use in ELISA and Flow Cytometry. Other applications not tested.
Recommended Dilutions:
Flow Cytometry: 5ug/ml (80ul)
Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol:
1. Ficoll prepared human peripheral blood mononuclear cells (PBMC) were stimulated overnight in culture with 5ug/ml PHA.
2. Five x10e5 cells/tube were incubated 45 minutes on ice with 80ul of C2549-22P1 at 5ug/ml.
3. Cells were washed twice and incubated with Goat anti-Mouse IgG/FITC, after which they were washed twice, fixed and analyzed by FACS using a lymphoid gate.
4. Cells stained positive with a mean shift of 0.33 log10 fluorescent units when compared to a Mouse IgG1 negative control at a similar concentration.
5. Binding was blocked when reagent was pre incubated with a 10-fold excess of C2549-22R.
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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