Hoefer SE400 - Manual (Page 42)

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Note: Because this is a highly
sensitive staining method, it
is important to wear gloves
when handling gels and to use
clean containers. To reduce the
background, use only high-purity
reagents and remove all buffer
from the gels during the fixing
and destaining steps.
Coomassie Stain Protocol
A. Stain gel in coomassie stain solution at room
temperature overnight. Gels can also be stained
rapidly by placing them at 55 °C in a shaking water
bath for 30– 45 min.
B. Place gel in Destain solution I at room tempera-
ture. Change the destain solution when it reaches
a deep blue color until clear background results.
C. Store the gel in Destain solution II.
For a more sensitive method, silver stain protocol is
recommended.
Silver Stain Protocol
(adapted from Morrissey, 1981)
Gentle agitation is recommended throughout
this procedure.
A. Stain the gel as usual with Coomassie Blue.
Destain the gel with several changes of
Destain solution II.
—OR—
Fix the gel in 100 ml Destain solution I for
30 minutes, then place the gel in 100–200 ml
Destain solution II for 30 minutes. Discard the
solution, refill, and wash with Destain solution II
a second 30 minutes.
B. Transfer the gel to 100 ml crosslinking solution
for 30 minutes.
C. Decant the glutaraldehyde and rinse the gel
with several changes of deionized water over a
period of two hours.
—OR—