Western blot and Triton X-100 in blocking buffer - (Feb/20/2013 )
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.
we used tween 20, not triton.