A Film Detection Method for Tritium‐Labelled Proteins and Nucleic Acids in Polyacrylamide Gels

FEBS Journal - Tập 46 Số 1 - Trang 83-88 - 1974
William M. Bonner1,2, Ronald A. Laskey2
1Department of Health, Education and Welfare, National Institutes of Health, 9000 Rockville Pike, Bethesda, Maryland, U.S.A. 20014
2M. R. C. Laboratory of Molecular Biology, Postgraduate Medical School, University of Cambridge, Hills Road, Cambridge, Great Britain CB2 2QH

Tóm tắt

A simple method is described for detecting 3H in polyacrylamide gels by scintillation autography (fluorography) using X‐ray film. The gel is dehydrated in dimethyl sulphoxide, soaked in a solution of 2,5‐diphenyloxazole (PPO) in dimethylsulphoxide, dried and exposed to RP Royal “X‐Omat” film at ‐70 °C. Optimal conditions for each step are described. β‐particles from 3H interact with the 2,5‐diphenyloxazole emitting light which causes local blackening of an X‐ray film. The image produced resembles that obtained by conventional autoradiography of isotopes with higher emission energies such as 14C. 3000 dis. 3H/min in a band in a gel can be detected in a 24‐h exposure. Similarly 500 dis./min can be detected in one week.When applied to the detection of 35S and 14C in polyacrylamide gels, this method is ten times more sensitive than conventional autoradiography. 130 dis. 35S or 14C/min in a band in a gel can be detected in 24 h.

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Tài liệu tham khảo

10.1038/182524a0

10.1016/0003-2697(70)90100-4

10.1038/227680a0

10.1073/pnas.69.12.3665

10.1021/bi00842a023

10.1016/0022-2836(70)90069-0

10.1016/S0021-9673(01)80075-3

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Thông tin xuất bản

FEBS Journal

Tập 46 Số 1

83-88

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