ISSN:
0173-0835
Keywords:
Chemistry
;
Biochemistry and Biotechnology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Chemistry and Pharmacology
Notes:
The 45Ca filtration method which allows calcium-binding proteins to be detected after their adsorption to nitrocellulose filters was used to test conditions for improved Western blotting of calmodulin. Our results indicate that dodecyl sulphate is not efficiently removed from calmodulin during electroelution in standard transfer buffers. This detergent disrupts binding of calmodulin to nitrocellulose. Electrotransfer in potassium buffer which precipitates dodecyl sulphate leads to a 6-fold increase in calmoculin retention by nitrocellulose. With this and other modifications, the detection limit of the 45Ca overlay method of Maruyama et al. (J. Biochem. 1984, 95, 511-519) for calmodulin is increased to 0.2 μg per lane.
Additional Material:
6 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/elps.1150090704
