||CAPN2 was purified by sequential chromatography through DEAE-Sepharose, A 1.5m Bio-Gel, and Phenyl-Sepharose CL-4B columns.
||50mM Imidazole-HCl, 0.2mM EDTA, 1mM DTT and 50% glycerol, pH 7.4.
||CAPN2 consists of an 80 kDa large subunit and a 30 kDa small subunit.
||Greater than 90% as determined by SDS-PAGE.
||his protein can be used for immunoblots, absorption experiments in immunohistochemistry, radioimmunoassay and intracellular injection. For adsorption we suggest the following procedure: A- Dilute 1μl of the antiserum against m-calpain in 1ml of the usual buffer for immunohistochemistry (final dilution 1:1,000). B- Add 1μg of protein to 1ml of the diluted antibody solution and mix well. C- Incubate for at least 6 hours in the cold. D- Apply to tissue-sections and incubate for 3 days. E- Complete the immunohistochemical reaction as usual (biotinylated second antibody, ABC-complex, DAB). As a result, the immunostaining should be strongly reduced or even completely prevented.
||CAPN2 although stable at 10°C for 1 week, should be stored desiccated below -18°C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Please prevent freeze-thaw cycles.