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Applications: Useful for screening for inhibitors of PD-1 binding to PD-L1
Contraindications: Only limited amounts of DMSO can be included, as it has been shownto disrupt PD-1-PD-L1 interaction. Avoid green and blue dyes that absorb light in theAlphaScreen signal emission range (520-620 nm), such as Trypan Blue. Avoid the use of thepotent singlet oxygen quenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+,Zn2+ and Ni2+). The presence of >1% RPMI 1640 culture medium leads to a signal reduction dueto the presence of excess biotin and iron in this medium. MEM, which lacks these components,does not affect AlphaScreen assays.
Description: The PD-1[Biotinylated]:PD-L1 Homogeneous Assay Kit is designed to measurethe inhibition of PD-1 binding to PD-L1. The PD-1[Biotinylated]:PD-L1 Homogeneous Assay Kitcomes in a convenient AlphaLISA® format with purified biotinylated PD-1, FLAG-tagged PD-L1,and assay buffer to perform a total of 384 reactions. With this kit, only three simple steps on amicrotiter plate are required. First, a sample containing biotinylated PD-1 and an inhibitor ofchoice is incubated with the FLAG-tagged PD-L1 for 60 minutes. Next, acceptor beads areadded, then donor beads, followed by reading the Alpha-counts.
Storage Stability: Stable at least 6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Lin, D., et al. Proc Natl Acad Sci U.S.A. 2008, 105: 3011-3016.
2. Keir, M.E. et al. Annu. Rev. Immunol. 2008, 26: 677-704.
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