Re: [ccp4bb] measure detergent concentration

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: measure detergent concentration
From: "Edward A {- dot -} Berry" BerryE {- at -} UPSTATE {- dot -} EDU
Date: 2009-10-23

Next message:
Subject: Re: measure detergent concentration
From: Michael Matho mmatho {- at -} SCRIPPS {- dot -} EDU
Date: 2009-10-23

Subject: Re: measure detergent concentration
From: Patrick Loll pat {- dot -} loll {- at -} DREXEL {- dot -} EDU
Date: 2009-10-23

I'll second this. We've done this as an exercise in NSLS Membrane
Protein Crystallization workshop for a few years, and it works like a
charm. You can stain in a warm iodine chamber and visualize by
scanning the TLC plate on a garden variety scanner (we use an
inexpensive Canon LIDE that probably cost less than USD 60 five years
ago). We quantify the spot intensity with NIH Image or equivalent, and
get lovely linearity down to the CMC, spotting only 1 uL of sample--so
we haven't seen any need to concentrate.

On 23 Oct 2009, at 3:41 PM, Edward A. Berry wrote:

> Only easy if you happen to have silica gel TLC plates and
> a chromatography jar lying around, perhaps from some
> phospholipid analysis:
>
> A strategy for identification and quantification of
> detergents frequently used in the purification of membrane proteins
> Laura R. Eriks, June A. Mayor, and Ronald S. Kaplan
> Analytical Biochemistry 323 (2003) 234–241
>
> This paper recommends spotting on a TLC plate and running
> beside standard amounts of the same detergent. From intensity/size
> of the detergent spot after developing you can bracket the detergent
> concentration. (And by the way they found that detergents are
> concentrated by ultrafiltration). To increase sensitivity, speedvac
> a volume too large to
> spot on the plate, dissolve the residue in Me0H.
>
> Ed
> weikai@CRYSTAL.HARVARD.EDU wrote:
>> Hi Folks:
>> After concentrating a membrane protein, is there a (easy) way of
>> measuring
>> the detergent concentration in the sample?
>> Regards,
>> Weikai

---------------------------------------------------------------------------------------
Patrick J. Loll, Ph. D.
Professor of Biochemistry & Molecular Biology
Director, Biochemistry Graduate Program
Drexel University College of Medicine
Room 10-102 New College Building
245 N. 15th St., Mailstop 497
Philadelphia, PA 19102-1192 USA

(215) 762-7706
pat.loll@drexelmed.edu

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: measure detergent concentration
From: "Edward A {- dot -} Berry" BerryE {- at -} UPSTATE {- dot -} EDU
Date: 2009-10-23

Next message:
Subject: Re: measure detergent concentration
From: Michael Matho mmatho {- at -} SCRIPPS {- dot -} EDU
Date: 2009-10-23