Protein determination

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JohnRowland
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Protein determination

Hi, does anyone know of an effective method for determining protein concentration in the presence of non-ionic detergents such as Triton X-100?

Any help would be greatly appreciated.

Soudabeh
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JohnRowland wrote:Hi, does

JohnRowland wrote:

Hi, does anyone know of an effective method for determining protein concentration in the presence of non-ionic detergents such as Triton X-100?

Any help would be greatly appreciated.

There is a BCA assay that you can use. In the manual, it will say what chemicals are compatible with this assay. However, as long as you make your standard in the same buffer as the protein, it will be fine because the non-ionic substance will be diluted out. And if it gives you a color in the background the standard will also have the same color.

jangy
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I used to make my own

I used to make my own standard curve and results on TLC plates stained with ninhydrin

nin1318
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pierce BCA kit works for this

pierce BCA kit works for this. they also have a reducing-agent compatible kit which allows for lots of other stuff to be in the sample and it still works (even BME).

bwbrian
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Here's a link to the Pierce

Here's a link to the Pierce BCA webpage. They claim you can have up to 5% Triton X-100 in your sample. There is also an instruction book you can link to near the bottom of the page.

http://www.piercenet.com/Objects/View.cfm?type=ProductFamily&ID=02020101