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Assay Method Information

Assay Name:  HTS for LYP Inhibitors-an Autoimmunity Target - Primary screen
Description:  LYP, a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling, is encoded by the PTPN22 gene. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave#s disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that its effect could be eliminated by a specific small-molecule inhibitor. Assay Principle. Full enzymatic activity is present in a 62 kDa N-terminal catalytic domain of the phosphatase that can be expressed in and purified from bacteria. Incubation of this recombinant protein with the appropriate substrate, DiFMUP (6,8-difluoro-4-methylumbelliferyl phosphate) results in the conversion of substrate to a fluorescent derivative. The endpoint of the reaction is assessed by measuring fluorescence with excitation at 360 nm and emission at
Affinity data for this assay
 

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Last update November 1, 2007
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