发信人: moso (野鬼), 信区: Biology
标 题: Re: about protein con. in SDS buffer
发信站: Unknown Space - 未名空间 (Mon Jun 13 16:12:04 2005), 转信

Try this:
Add x ul of your sample to in total 200ul water(remember to do your standard
curve as the same way using BSA)
Add 10ul of 250mM CaCl2 and 25ul of 200mM Na3PO4 (pH 7.4) to co-precipitate
protein at room temperature for 10min
Add 1ml ethanol, mix well to wash the precipitate
Centrifuge at 7000 x g for 1min and aspirate supernatant.
Add 100ul water and 1ml ethanol to wash the pellet again
Centrifuge as above to remove supernatant
To the pellet, add 100ul 5 X Bradford solution
Incubate at room temperature for 15min
Add 400ul water to the solution
Read it at A595

It works for me very well. Concentration can range from 1-20ug/ul
【 在 dh (dh) 的大作中提到: 】
: Does anyone knows how to measure protein concentration in SDS loading buffer?
: I tried the bio-rad protein assay reagent but it only work without detergent
: and reducing agent. your help will be really appreciated.


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