Hello...problem of detecting proteins with CBB staining

Daniel Prieto via methods%40net.bio.net (by dprieto from fcien.edu.uy)
Mon Jul 30 14:45:28 EST 2012


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: 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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