Non disponible en dehors du Royaume-Uni et de l'Irlande

Application

Calpain inhibitor I inhibits calpain, which activates myosin light chain kinase and protein kinase C by partial proteolysis. The inhibitor should be membrane permeable due to low molecular weight and lack of charged residues.Calpain Inhibitor I is used for western blotting methods.

Formula variant

C₂₀H₃₇N₃O₄

General description

Calpain Inhibitor I is a strong, competitive inhibitor of the Ca²⁺-dependent neutral cysteine proteases calpain I and calpain II.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Preparation Note

Working concentration: 0.1 - 10 µMThe suggested starting concentration is 17 µg/ml. This is the concentration at which half maximal inhibition of calpain I is observed.Working solution: Recommended solvent is DMF, ethanol, or methanol up to 10 mg/ml.Storage conditions (working solution): -15 to -25 °CSolutions in DMF, methanol, ethanol are stable for 4 weeks at -15 to -25 °C.

Quality

Purity: 98% (from C); chromatographically homogeneous

Reconstitution

Soluble in DMF, ethanol, or methanol to 10 mg/ml. For a stock solution, Roche recommends dissolving 1 mg of the inhibitor in 100 µl DMF, methanol, or ethanol. Before use, dilute with water or phosphate buffer (0.1 M, pH 7.5) to desired concentration. Solutions in DMF, ethanol, or methanol are stable for 2 to 3 days at 2 to 8 °C, and approximately 4 weeks at -15 to -25 °C. For best results, prepare solutions fresh before use.

Specificity

Calpain Inhibitor I is a strong, competitive inhibitor of the Ca²⁺-dependent neutral cysteine proteases calpain I (requiring 2–75 µM Ca²⁺ for activation) and calpain II (requiring 200–800 µM) with a K_i-value between 0.12 µM and 0.23 µM. Calpain II is inhibited to a lesser extent than calpain I. Calpain Inhibitor I is also a strong inhibitor of papain and cathepsin B and L. It shows only weak inhibition of cathepsin H and α-chymotrypsin and does not inhibit trypsin. ID₅₀ for 0.02 U platelet calpain is 0.05 µM.