Jacob,
Coomassie fluorescence is linear over a wide range of concentrations
(citation below). You need access to an infrared scanner in order to
use this technique. I've used a Li-cor Odyssey scanner with good
results (http://www.licor.com/bio/odyssey/index.jsp).
Anal Biochem. 2006 Mar 15;350(2):233-8.
Quantitation of protein on gels and blots by infrared fluorescence of
Coomassie blue and Fast Green.
Luo S, Wehr NB, Levine RL.
Coomassie blue staining of gels and blots is commonly employed for
detection and quantitation of proteins by densitometry. We found that
Coomassie blue or Fast Green FCF bound to protein fluoresces in the
near infrared. We took advantage of this property to develop a rapid
and sensitive method for detection and quantitation of proteins in
gels and on blots. The fluorescence response is quantitative for
protein content between 10 ng and 20 microg per band or spot. Staining
and destaining require only 30 min, and the method is compatible with
subsequent immunodetection.
PMID: 16336940
Regards,
Steve
--
Steven Darnell
University of Wisconsin-Madison
Madison, WI USA
Jacob Keller said the following, on 09/25/2008 04:35 PM:
Dear Crystallographers,
I remember having seen on this listserve that coomassie stain is
horribly non-linear in intensity per protein concentration, which
leads me to two questions:
1. Does anybody have a reference for quantitation of coomassie's
linearity (and possibly other stains), i.e. where and how extensive
the linear range is, and
2. Can anybody suggest a more linear protein gel stain?
Thanks,
Jacob Keller
*******************************************
Jacob Pearson Keller
Northwestern University
Medical Scientist Training Program
Dallos Laboratory
F. Searle 1-240
2240 Campus Drive
Evanston IL 60208
lab: 847.491.2438
cel: 773.608.9185
email: [EMAIL PROTECTED]
*******************************************
