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Is there a protocol for this product (target concentration, pH range for fluorophore, excitation and emission wavelengths, etc.)? Also, is this fluorophore quenched by the peptide and fluorescent only when cleaved?
Asked by: Ray_B
sc-300159 is a fluorogenic substrate for caspase-4 and caspase-1 (kcat/Km = 2.41 x 105 M-1s-1). Ac-WVAD-AMC exhibits approximately 2 fold greater affinity towards caspase-1 than for caspase-4.
Fluorescence is seen after cleavage. In assay the AMC liberated from the fluorogenic substrate was measured in a spectrofluorometer with an excitation wavelength of 380 nm and emission wavelength range of 430-460 nm.
Reconstitute sc-300159 before use in 1 ml DMSO for a concentration of 1mg/mg in DMSO. Store the reconstituted substrate at -20°C for up to 1-2 months and avoid repeated freeze-thaw cycles.
D. Lee, et al., Bioorg. Med. Chem. Lett. 9, 1667 (1999)
T.A. Rano, et al., Chem. Biol. 4, 149 (1997)
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