Hi,
Someone please explain me the exact solutions which can stain and destain the gels after SDS-PAGE.
The choice of the best method depends, to a large extent, on the required sensitivity. Coomassie Blue is the most commonly used stain. If greater sensitivity is needed, silver stain or a fluorescent stain can be used instead.
Look in a textbook or on google for coomassie blue or Ponceau Red or silver nitrate stainings. These three are the classical ones.
The choice of the best method depends, to a large extent, on the required sensitivity. Coomassie Blue is the most commonly used stain. If greater sensitivity is needed, silver stain or a fluorescent stain can be used instead.
- For staining coomassie blue in acetic acid methanil
- For destaining agitate gel at room temp for 30 min to over night also in acetic acid methanol
- initially cover gel in acetic acid methanol then gently rock. Pour off when mixture has turned blue ( leached coomassie blue) after about 10 min. Add fresh. Incubate for a further 30 min. Most gels will be destained removing background using this method in 1-2 hours
Dear
1-For staining follow this procedure
- Fix gel in Fixing solution for 1 hr to overnight with gentle agitation. Change
solution once at first 1 hr.
- Stain gel in Staining solution for 20 min with gentle agitation.
- Destain gel in Destaining solution. Replenish the solution several times until
background of the gel is fully destained.
-Store the destained gel in Storage solution
2-Method for destaining
Follow experimental design of G. Hervieu. 1997
Regards
http://ipmb.sinica.edu.tw/proteomics/Documents/Coomassie%20Staining.pdf
http://www.sciencedirect.com/science/article/pii/S1366212008700485
If the protein concentration is sufficiently high, go for CBB staining method or else go for silver staining technique.
I guess it will depend also in what do you want to do with the protein after running it in SDS-PAGE as some stains are not compatible with subsequent assays. Choose wisely
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