Affinity chromat.

Product: 6-His human calretinin (recombinant) produced in Escherichia Coli.Lot No.: 22Form: lyophilized (10 or 100 µg in 25, resp. 155 µl of: 10 mM Tris, 50 mM NaCl, 2 mMCaCl2, 1 mM EDTA, 1mM ß-mercaptoethanol, pH 7.4).DescriptionThis recombinant protein has been purified by affinity chromatography on a NTA-resinThis protein can be used for immunoblots, absorption experiments in immunohistochemistry,radioimmunoassay and intracellular injection.For adsorption we suggest the following procedure:- Dilute 1 µl of the antiserum 7696 (SWant) against calretinin in 5 ml of the usual buffer forimmunohistochemistry (final dilution 1:5'000).- Add 1 µg of the recombinant protein to 1 ml of the diluted antibody solution and mix well.- Incubate for at least 6 hours in the cold.- Apply to tissue-sections and incubate for 3 days.- 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.StorageFreeze in small aliquots (e.g. 1 or 10 µg) and keep at -80°C. Avoid repeated freezing andthawing.ReferencesSchwaller B., Buchwald P., Blümcke I., Celio M.R. and Hunziker W. (1993) Characterizationof a polyclonal antiserum against the purified human recombinant calcium-binding proteincalretinin. Cell Calcium 14: 639-648.