Re: [ccp4bb] Copper Staining

Protein crystallography Programs for crystallography X-ray detectors
Basic tutorials:
Protein Peptide

CCP4bb navigation

CCP4bb <-- 2008 <-- March 2008 <-- 03 March 2008

Previous message:
Subject: Re: Copper Staining
From: Jacob Keller j-keller2 {- at -} MD {- dot -} NORTHWESTERN {- dot -} EDU
Date: 2008-03-03

Next message:
Subject: superimpose two maps
From: Yanming Zhang yzhang {- at -} MAILER {- dot -} SB {- dot -} FSU {- dot -} EDU
Date: 2008-03-03

Subject: Re: Copper Staining
From: Dima Klenchin klenchin {- at -} FACSTAFF {- dot -} WISC {- dot -} EDU
Date: 2008-03-03

>There's an even faster reagent for staining - takes abut 1 minute overall
>and the results look just like dear old Coomassie. This reagent works
>wonderfully for us. Yes, the recipe is proprietary - but if you add up the
>cost of reagents for normal Coomassie and compare - this one isn't much more
>expensive.
>
>http://www.novexin.com/products/InstantBlue.html

Sounds like a good old Bradford reagent AKA colloidal Coomassie G-250.
60 mg G-250 dissolved in 50 ml ethanol and diluted into 1 L of
1/10 dilution of concentrated phosphoric acid (~8.5% final).
"Solubilizing agents" is a smoke screen, for G-250 is fully soluble
in ethanol and for the colloid particles to not penetrate the gel
pores inclusion of any solubilizing agents will only be detrimental.

Betcha it's a browish solution that forms blue pellet when
centrifuged hard.

If so, the claimed 5-10 ng sensitivity can only be achieved
after 1) at least 15 min staining (30 min better; no one abolished
laws of physics that requre diffusion into the gel for the
interaction to take place), 2) some destaining to get rid of
slight brown-blue background.

Dima

CCP4bb navigation

CCP4bb <-- 2008 <-- March 2008 <-- 03 March 2008

Previous message:
Subject: Re: Copper Staining
From: Jacob Keller j-keller2 {- at -} MD {- dot -} NORTHWESTERN {- dot -} EDU
Date: 2008-03-03

Next message:
Subject: superimpose two maps
From: Yanming Zhang yzhang {- at -} MAILER {- dot -} SB {- dot -} FSU {- dot -} EDU
Date: 2008-03-03