On 26 Sep 2008, at 08:20, Jacob Keller wrote:
> 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.
You can visualize Sypro Orange on a regular Ethidium bromide detection
system, but a computer based detection system is more sensitive.
Jürgen
>
>
> 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: [log in to unmask]
> *******************************************
>
> ----- Original Message ----- From: "Gregory Alushin" <[log in to unmask]
> >
> To: <[log in to unmask]>
> 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: [log in to unmask]
>>> *******************************************
-
Jürgen Bosch
University of Washington
Dept. of Biochemistry, K-426
1705 NE Pacific Street
Seattle, WA 98195
Box 357742
Phone: +1-206-616-4510
FAX: +1-206-685-7002
Web: http://faculty.washington.edu/jbosch
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