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Re: [ccp4bb] Copper Staining |
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- Protein crystallographyMain steps:- Protein purification- Crystallisation Special:- Programs for crystallography- X-ray detectors Basic tutorials:- Chemistry- Protein - Peptide - Amino Acids Xtal community:- CCP4BB |
CCP4bb navigationCCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999Subject: 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 navigationCCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999 |
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