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Visualization Notes
Hoefer SE 1200 Easy Breeze Air Gel Dryer
13
Visualization Notes
Direct Autoradiography
Prepare the gel in the usual manner and expose to film as directed with the instructions
accompanying the film. Exposure times, especially for medium energy beta-emitters
such as
14
C and
35
S can be quite long. For maximum sensitivity of these types of labels,
use fluorography.
Fluorography
Fluorography intensifies a weak beta emission signal by impregnating the gel with a
suitable fluor that converts beta emission from
35
S,
14
C, and
3
H into light. Drying the
gel in cellophane does not quench this signal, but for
3
H, a very weak beta-emitter, up to
20% of the signal may be lost.
If using EN
3
HANCE or sodium salicylate as fluorography agents, include 0.5% to 2%
glycerol in the cold water treatment prior to drying. Dehydration steps are not required
for thinner, lower percentage gels, which are far less prone to cracking. If exposed to the
maximum drying temperature (≈54 °C), these gels may form longer fluor crystals,
resulting in higher background on X-ray exposures.
NOTE Certain fluorographic agent manufacturers specify temperatures higher than
those reached in the Hoefer Easy Breeze (e.g. Autofluor™ [National
Diagnostics] and Fluoro-Hance™ [Research Products International], which
require up to 80 °C to produce an optimum signal). This dryer is not
recommended for such agents.
Bibliography
Juang, R.H., Chang, Y.D., Sung, H.Y., and Su, J.C., Oven-drying method for
polyacrylamide gel slab packed in a cellophane sandwich. Anal. Biochem. 141, 348–350
(1984).
Laskey, R.A., The use of intensifying screens or organic scintillators for visualizing
radioactive molecules resolved by gel electrophoresis. (In Methods in Enzymology, 65,
L. Grossman and K. Moldave, eds.) Academic Press, N.Y. 365–371 (1980).
Perng, G.S., Rulli, R.D., Wilson, D.L. and Perry, G.W., A comparison of fluorographic
methods of detection of 35S labeled proteins in polyacrylamide gels. Anal. Biochem.
173, 387–392 (1988).