Well, that separation is not exactly correct. There are mixed procedures
where you stain with a mixture of both CBBs and some of them with the
former and some others with the latter. I would suggest you to take a look
at Candiano et al. 'Blue silver' (colloidal CBBG) protocol which works
nicely, w/o
background staining, fast and very sensitive (close to silver methods) and
compatible with MS.
All the best,
Daniel
On Mon, 30 Jul 2012, Jayakumar, R wrote:
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: [email protected]
[mailto:[email protected]] On Behalf Of sudheer sangeetham
Sent: Monday, July 30, 2012 4:40 AM
To: [email protected]
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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