Hi All,
thanks for the responses. It seems that coomassie linearity is not so bad
after all, especially when measured by IR fluorescence (reference from Steve
Darnell). Others mentioned silver stain, but it seems that silver has a
narrower dynamic range, even though more sensitive. Sypro and Biorad
flamingo seem linear, and as sensitive as silver. But...for the fluorescent
dyes, one optimally needs a fluorescence scanner.
Thanks all,
Jacob Keller
ps The original coomassie reference seems to be
S. Fazekas de St. Groth, R.G. Webster and A. Datyner, Two new staining
procedures for quantitative estimation of proteins on electrophoretic
strips, Biochim. Biophys. Acta 71 (1963), pp. 377-391.
*******************************************
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]
*******************************************
----- Original Message -----
From: "Gregory Alushin" <[EMAIL PROTECTED]>
To: <CCP4BB@JISCMAIL.AC.UK>
Sent: Thursday, September 25, 2008 6:07 PM
Subject: Re: [ccp4bb] Off-topic: coomassie linearity?
The Bio-Rad Flamingo Fluorescent Gel Stain has a very linear profile with
protein concentration (if you believe the standard curves in the manual).
In my hands it gives nice results for binding assays.
Cheers,
-Greg Alushin
On Sep 25, 2008, at 2:35 PM, Jacob Keller wrote:
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]
*******************************************
