Diagnostic & Veterinary
Caspase-8 IETD-R110 Fluorometric HTS Assay Kit
Descrition & Target
Caspase-8 IETD-R110 Fluorometric HTS Assay Kit is specifically designed for HTS-based assays. The kit provides a homogenous assay system for fast and highly sensitive detection of caspase-8 activity by fluorescence in enzymatic reaction or mammalian cells. The fluorogenic substrate (Ac-IETD)<sub>2</sub>-R110 contains two IETD tetrapeptides and is completely hydrolyzed by the enzyme in two successive steps. Cleavage of the first IETD peptide results in the monopeptide Ac-IETD-R110 intermediate, which has absorption and emission wavelengths similar to those of R110 (Î»<sub>Ex</sub>/Î»<sub>Em</sub> = 496/520 nm) but has only about 10% of the fluorescence of the latter. Hydrolysis of the second IETD peptide releases the dye R110, leading to a substantial fluorescence increase.
<p class="style5 style11"><strong>Note:</strong> While caspase-8 preferentially cleaves the consensus sequence IETD compared to other substrate sequences, other caspases such as caspase-3 also can cleave IETD efficiently. Overlapping caspase substrate recognition limits the usefulness of caspase substrate peptides for distinguishing between different caspase activities in cell lysates.</p>
The assay kit includes Ac-IETD-CHO, which is a caspase-8 inhibitor and can be used as a negative control. Also, R110 is provided in the kit for generating a standard curve, which can be used for quantifying caspase-8 activity.
<p class="style5 style11"><strong>Kit Components: </strong></p>
<li>Cell lysis/assay buffer</li>
<li>Enzyme substrate (Ac-IETD)2-R110</li>
<li>Enzyme inhibitor Ac-IETD-CHO</li>
<p class="style5 style11"><strong>Features:</strong></p>
<li><b>HTS-compatible:</b> Single-step homogenous assay specifically designed for HTS-based detection.</li>
<li><b>Fast: </b>Fast enzyme kinetics.</li>
<li><b>Sensitive:</b> The enzymatic reaction forms intensely green fluorescent rhodamine 110 (R110) product.</li>
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