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Calpain Assay Kit

Components:

10x Cell Lysis Solution- 25 ml, store at 4ºC
10x Calpain Buffer- 0.5 ml each, store at -20ºC
Casein Solution- 1 ml, store at 4ºC
Calpain Assay Solution- 20 ml, store at 4ºC (for 200 assays)

Product Description: Calpains are a family of cysteine proteases involved in the regulation of many protein substrates via limited proteolysis in response to calcium signaling. This feature of calpains notably differs from proteasomes and autophagy, both of which degrade their protein substrates. Two well-characterized members of the conventional calpain family in mammals are calpain 1 (mu isoform) and 2 (m isoform), which are heterodimers of a specific 80-kDa large subunit and a common 28-kDa small subunit. Calpains have been implicated in numerous calcium-regulated processes such as cell motility, cell cycle progression, cell death, and differentiation, making them ideal therapeutic targets. The Calpain Assay Kit is derived from a modified colorimetric method (Arch Biochem Biophys 319:211,1995) for rapid analysis of calpain activity in crude lysates. Inclusion of 5 uM or 5 mM CaCl2 in the assay buffer allows one to determine the activity of calpain 1 and calpain 2, respectively. Casein is used as substrate in the assay kit and its breakdown is measured by a simple protein-dye binding method. The signal is proportional to the amount of total calpain activity. Note that dithiothreitol (DTT) is required for preparation of crude lysates, but is not included in the kit. The kit is stable for at least one year under proper storage and handling conditions.