Caspase 3 Activity Assay Kit

Product Description

The Caspase 3 Activity Assay Kit utilizes spectrophotometry to detect the activity of caspase 3 enzyme in cell or tissue lysates, or purified caspase 3 enzyme.

Caspases (Cysteine-requiring Aspartate Proteases) are a family of proteases that play crucial roles in the process of apoptosis. Caspase 3, also known as CPP32, Yama, or apopain, and sometimes written as caspase-3 or caspase 3, belongs to the CED-3 subfamily of the caspase family. It is a key enzyme in the process of apoptosis. Caspase 3 is one of the most studied caspases in mammalian cells. It can cleave procaspases 2, 6, 7, and 9, and can directly cleave many caspase substrates, including PARP (poly(ADP-ribose) polymerase), ICAD (Inhibitor of caspase-activated deoxyribonuclease), gelsolin, and fodrin, among others. These protein cleavages mediated by caspase 3 are integral parts of the molecular mechanisms of apoptosis. Additionally, Caspase 3 also plays a critical role in nuclear apoptosis processes, including chromatin condensation, DNA fragmentation, etc. Furthermore, caspase 3 is crucial for cell blebbing.

This assay kit is based on the ability of Caspase 3 to catalyze the substrate Ac-DEVD-pNA to produce yellow pNA (p-nitroaniline), which can be detected by measuring absorbance. pNA exhibits strong absorption near 405nm.

When detected using a microplate reader or spectrophotometer with cuvettes that can hold up to 100μl, in addition to the standard curve, the kit can detect 20 samples (20T) or 100 samples (100T).

Performance

HL60 cell caspase3 activity assay

The figure shows that HL60 cells were divided into non-dosing group (Con) and apoptosis-induced group (Apo), and were incubated with substrate for 0h, 2h, 4h, 6h, 8h to overnight using MeilunBio and other international brands of Caspase3/7 activity assay kits, and the results were recorded at A405nm on an enzyme-labeled instrument, respectively. The red line in the figure represents Apo group, the blue line represents Con group, and the green line represents blank group.

From the experimental results, the detection sensitivity of Apo group was better than that of international brand, and the A405 reading of apoptosis-induced group was about 3 times of that of Con group; while the international brand was only 2 times and the reading was lower than that of Apo group.

Shipping and Storage 

• Storage：Store at -20°C. Ac-DEVD-pNA and pNA should be protected from light.
• Shipment：

Usage Statement

Research Use Only (RUO)