This reagent is intended for research purposes only. The Caspase-3 kit is a solid-phase sandwich ELISA designed to quantify caspase-3 levels in biological samples. The procedure involves adding both standard solutions with known caspase-3 concentrations and unknown samples into microplate wells pre-coated with capture antibodies. Following an incubation period, biotinylated detection antibodies are introduced, allowing them to bind to the captured caspase-3. After thorough washing to remove unbound components, avidin-conjugated HRP (horseradish peroxidase) is added. This step enables the formation of a stable antibody-enzyme complex through the interaction between biotin and avidin. Another incubation and wash cycle ensures that only the bound enzyme conjugate remains. Subsequently, substrates A and B are added simultaneously, triggering a colorimetric reaction. The intensity of the resulting color is directly proportional to the amount of caspase-3 present in the sample. This method provides a sensitive and specific way to measure caspase-3 activity, making it ideal for apoptosis studies and related research applications. The entire process is highly reproducible and suitable for high-throughput screening when performed according to the manufacturer’s instructions.
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