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cell lysate for pulldown


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3 replies to this topic

#1 soymilk14

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Posted 10 November 2010 - 09:56 PM

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

#2 madrius1

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Posted 12 November 2010 - 09:43 AM

Fresh is always better.

#3 madrius1

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Posted 12 November 2010 - 09:43 AM

Fresh is always better.

#4 shivasankari

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Posted 18 November 2010 - 06:31 AM

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
Cheers

Shiva




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