ISSN:
0173-0835
Keywords:
Chemistry
;
Biochemistry and Biotechnology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Chemistry and Pharmacology
Notes:
Film detection of compounds labeled with the weak beta-emitters 3H and 14C is demonstrated for electroimmunoprecipitated proteins in agarose gels by means of the water-soluble fluorophor sodium salicylate. The degree of enhancement of film detection of tritium and carbon-14 with 0.7 M sodium salicylate paralleled fluorography with diphenyloxazole on polyacrylamide gels. The method was optimized and the sensitivity, expressed as radioactivity per cm immunoprecipitate, was 0.9 nCi/cm (2000 dpm/cm) in 24 h for 3H and 0.4 nCi/cm (830 dpm/cm) in 18 h for 14C. The procedure was found superior to the commercially available enhancement solutions ‘EnHance’ and ‘Amplify’. Salicylate fluorography on Kodak X-Omat L and AR-5 films was about 30 and 60 times more sensitive than Ultrofilm 3H. With 14C-labeled anti-antibodies rocket electroimmunoprecipitation corresponding to about 50-100 pg protein could be visualized in 7 days. Conclusively, the method is rapid (soaking and drying require about 30 min), sensitive, reproducible, cheap, and advantageous if the immunoplates are to be stained for protein after fluorography.
Additional Material:
7 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/elps.1150050503
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