Protein silver staining.

David Martin damartin at bioslave.uio.no
Wed Mar 6 05:41:44 EST 1996


In article <4hieuh$2248 at yuma.ACNS.ColoState.EDU>,
   "Stephen R. Decker" <Sdecker at Vines.colostate.edu> wrote:
-->Belinda,

<snip out Ag stain protocol>
-->
-->If yo are in a hurry to get the gel, you can cut the first two steps back 
to 15 mins 
-->each and the long wash to about an hour, if you don't mind a little 
background.
-->

If you are in a _real_ hurry and want a result in 10 mins (and don't mind a 
bit of background) then you can speed things up a little:
Use a clean glass lasanga dish about 8-9 inches (20-25cm) square

Place gel on the dish without touching it (split the plates and drop the gel 
carefully)
1. 10% Acetic acid 40% Methanol, 100 ml, 1 min full power in the microwave.
2. Pour off. rinse briefly with dH20 (30 secs).
3. Pour off. add 100ml dH20 containing 100ul 5mg/ml DTT. Microwave 1 min.
4. Pour off. add 100ml 0.1% AgNO3. Microwave 1 min. agitate 2 min.
5. Pour off. Add 100 ml developer (3% Na2CO3 0.04% formaldehyde), swirl till 
the liquid is brown then pour off.
6. Add 100ml developer and develop the bands.
7. stop by addition of citric acid

Its most definitely not the prettiest silver stain but if you want to cast, 
run, and stain a mini gel in 90 mins........

....d

=========================================================
* David Martin, PhD -  Post-Doctoral Research Fellow    *
* Atherosclerosis and Thrombosis research group         *
* Biotechnologisenteret i Oslo                          *
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