cell lysate for pulldown - (Nov/10/2010 )

hello everyone,

i would like to detect the gtp bound form of a protein from my gst pulldown. my question is: should i always use fresh cell lysate or i can stock my cell lysate and perform pulldown?

thank you so much

Fresh is always better.

Fresh is always better.

Hi,

Storing the lysate is a very bad idea, cos it contains proteases and the proteases generally cleave your protein of interest.So, you tend to get different results each time. You can store the cells, not the lysate!

Shiva