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Harvesting cells


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#1 Biochem_newbie

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Posted 24 September 2014 - 06:37 AM

Hi there, 

 

I was wondering if someone can confirm if this is right method or wrong method for harvesting cells. 

 

For 293s, resuspend the cells with cold PBS, spin down, discard the sup, and add lysis buffer with protease inhibitor dissolved. 

 

Is this correct?



#2 Tabaluga

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Posted 24 September 2014 - 07:02 AM

So you wanna harvest cells - but what for ?


Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 


#3 Biochem_newbie

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Posted 24 September 2014 - 07:45 AM

Apologies for the vagueness.

I would like to prepare over-expressed samples in 293 for WB



#4 bob1

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Posted 25 September 2014 - 08:09 PM

In that case, yes, that is one method of preparing lysates. There are a number of other options as well, but basically they all involve washing the cells in a buffer then lysing.




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