Hello...problem of detecting proteins with CBB staining

Jayakumar, R via methods%40net.bio.net (by R.Jayakumar from roswellpark.org)
Mon Jul 30 13:52:14 EST 2012


Perhaps you should put that question to the author.  I have never felt the need to heat up a gel to stain it with CBB.   Silver staining is still the most sensitive method to detect proteins, and if it si working only intermittently, perhaps you should look at your handling procedures.   
   And secondly what kind of CBB dye are you using, the R-250 or the G-250?.  Remember only the former is used for protein staining while the later is used to estimate proteins.
Best of luck
Jay


-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu [mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of sudheer sangeetham
Sent: Monday, July 30, 2012 4:40 AM
To: Methods from magpie.bio.indiana.edu
Subject: Hello...problem of detecting proteins with CBB staining

Hello
I followed the method according to this article i.e Heating Greatly Speeds Coomassie Blue Staining and Destaining BioTechniques 28:426-432 (March 2000). In that article they have shown they could able to detect up to 10ng of protein, which is clearly visible on the gel. * *I followed exactly similar conditions but I could able to detect 60ng of protein.
Actually I am looking for most sensitive method to detect protein on PAGE.
I tried manual Silver staining method(Blue et al) , some times it is working sometimes not. Could anyone please help me out with the problem. If anyone suggest the method, which is most sensitive and fast doing for detecting proteins that would be more help for me.

Thank you in advance.

--
Sudheer Babu.S
Research Fellow
Institute of Biochemistry
Biological Research Center
Szeged,Hungary.
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