Coomassie Blue: destaining

Allan Stensballe allan at NUKESPAMMERSstensballe.com
Sat Jul 15 14:01:35 EST 2000


On Fri, 14 Jul 2000 11:41:17 +0100, nospam at our.site wrote:

>This message has been posted by:  "Luis M. Martins" <L.Martins at REMOVE-THIS-TO-SENDicrf.icnet.uk>
>
>Hi 
>
>I am using PhastGel BlueR coomassie blue dye from Pharmacia, I am concearned
>with destaining the gel without losing the signal from the bands (I have
>around 10ng protein per band)
>
>I am currently destaining in 10% Acetic acid, this takes quite a long time
>and leaves an uneven blue background. I was considering using methanol to
>improve solubility of the CB stain. How much methanol should I use without
>risking loosing signal from the bands?
>
>Your help is much appreciated!
>
>Regards,
>
>Miguel


Hi Miguel & Group...

For visualization of ng levels of protein you should go for silver
staining instead of CB or Sypro dyes (assumed that you stain
electrophoretic gels and just need visualization).    
Silver staining with the right shortcuts is nearly as fast as CB, but
have sub ng sensitivity.

Staining techniques have recently been reviewed by Thierry Rabilloud
(Anal. Chem. jan. 1 2000, pp. 48A-55A)

Protocols for various staining techniques are reviewed at
www.protana.com (They are as good as the techniques allow and
compatible with mass spectrometry.)
See http://www.protana.com/services/protocols/default.asp

----------------------------------------------------
Allan Stensballe,
Protein Research Group,
University of Southern Denmark,
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