protein purification - (Oct/12/2008 )

Hi,

I want to purify protein from mouse tissue. I want to do it by using a piece of tissue from the ear.
did someone did something like this or have any suggestion

thank you

huh???

a specific protein?

in what abundance?

what is your plan for purification?

how much tissue weight will you start with?

or are you just trying to make a protein extract?

of cytosolic or membrane proteins?

soluble proteins?

your question can't be answered without more information.

I want to purify whole cell proteins to run western and to check a specific expresion of protein .

I don't know how much tissue I will start with, this is why I need help :-).
I want to have enough protein to run 30ug whole cell protein on western


I hope I made my self clearer

thank you for you help

for whole cell protein you can homogenize a small piece of tissue directly into gel loading buffer (without the marker dye (bromphenol blue) if you want to determine the protein concentration before loading) and heat. then centrifuge to eliminate insolubles.