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Western blot and Triton X-100 in blocking buffer


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#1 James.

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Posted 20 February 2013 - 05:19 AM

Hello,

I was wondering about the precise reason for using Triton X-100 in blocking buffer during a western blot. The cells containing my protein of interest are already lysed and triton x-100 is a detergent primarily used for permeabilizing cell membranes. However, I found a possible reason mentioned under the product's description at http://www.mclab.com/Triton-x-100.html

It says: 'It can be used to reduce the surface tension of aqueous solutions during immunostaining (usually in concentration of 0.1-0.5% in TBS or PBS Buffer).'

Could this be the reason for using triton x-100 in the blocking buffer? And can someone explain how exactly reducing surface tension would help in needing less antibody and getting a better surface covering? (I found this over at wikipedia, in the article about immunohistochemistry).

I thought reducing the surface tension could help in solubilizing the antibodies, allowing better contact and spreading of the antibodies, but I'm not sure.

Thanks in advance.

#2 Curtis

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Posted 20 February 2013 - 08:40 AM

we used tween 20, not triton.




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