Dr. David Martin
david.martin at read.my.sig
Tue Dec 17 07:14:47 EST 1996
In article <595too$12b at mserv1.dl.ac.uk>,
Dr Alok Adholaya <aloka at teri.ernet.in> wrote:
+>I am in a look out for a protocol for silver staining of proteins
+>seperated on SDS-PAGE. The protocol should have sensitivity of
+>conventional silver staining yet be rapid ( should not take more
+>than 15 minutes ). Does anyone know of such a procedure ??????
The extremely rapid silver stain follows: For best results use
spotlessly clean glass dishes/plates and do not touch the gel at all.
It is not as good a stain as taking several hours to do it properly
(background can be high) but it is sensitive and it does work.
Method: Very Rapid Silver Stain
Chemicals required: Coomassie Destain (40% MeOH 10% AcOH)
5 mg/ml Dithiothreitol
1 mg/ml AgNO3 10X
2.3M Citric Acid
Health and Safety: 40% Formaldehyde. Toxic in contact with skin.
1. Microwave gel for 1 minute at full power in 100 ml
2. Wash briefly in dH2O to re hydrate the gel (5-10 mins or
1min in microwave).
3. Microwave gel for 1 minute at full power in 100 ml dH2O
containing 100ul 5mg/ml DTT.
4. Wash in 0.1 mg/ml AgNO3 for 45 minutes to overnight.
5. Wash briefly in dH2O.
6. Wash in developer (40 ml 15% Na2CO3 200 ul 40% Formaldehyde
in 200 ml) till solution goes brown (100 ml for approx 1 minute).
7. Changge developer and develop. Stop by adding 5ml 2.3M
* David Martin - Atherosclerosis and Thrombosis research group *
* http://www.uio.no/~damartin/ david.martin at biotek.uio.no *
* Lab +47 22 95 84 54 Fax +47 22 69 41 30 GSM +47 90 74 27 65 *
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