Quantifying PLCγ2 Activity with C16CF3-coumarin

For the PLCγ2 assay with C16CF3-coumarin, mixtures of 0–40 nM of PLCγ2 and 10 μM of C16CF3-coumarin were prepared with micelle assay buffer. PLCγ2 (10 μl) and C16CF3-coumarin (10 μl) were dispensed into the microtiter plate, and the final concentrations were 0–20 nM and 5 μM, respectively. The fluorescence intensities from the assay reactions were monitored, as mentioned above. The slope was analyzed from the initial linear range (60 min) of reaction curves.

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Visvanathan R., Utsuki T., Beck D.E., Clayton W.B., Lendy E., Sun K.L., Liu Y., Hering K.W., Mesecar A., Zhang Z.Y, & Putt K.S. (2024). A novel micellular fluorogenic substrate for quantitating the activity of 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma (PLCγ) enzymes. PLOS ONE, 19(3), e0299541.

Publication 2024

Corresponding Organization : Cayman Chemical (United States)

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Variable analysis

independent variables

Concentration of PLCγ2 (0-20 nM)

dependent variables

Fluorescence intensity of the assay reactions

control variables

Concentration of C16CF3-coumarin (5 μM)
Micelle assay buffer

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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