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Vortexing agarose beads

immunoprecipitation

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#1 Artemis2007

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Posted 03 October 2014 - 01:43 PM

I have a couple of IP experiments using different kinds of agarose beads (protein A/G and strepavidin). Is it ok to vortex the beads to resuspend during the many washing steps? I understand the beads can be deformed by excess g forces, but I have no idea what level of g force is generated during a moderately fast vortex (I have found the very slow vortex not to be useful for this purpose).



#2 bob1

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Posted 04 October 2014 - 08:45 PM

Basically no, as any complexes formed during the ip could easily be separated by the vortexing. It's not the greatest forces but rather the swirling fluid that is the problem here.

#3 Artemis2007

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Posted 07 October 2014 - 11:55 AM

Thank you. I've been re-suspending by finger-flicking the eppendorf.  Wouldn't this also cause fluid swirling...?



#4 bob1

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Posted 09 October 2014 - 05:22 PM

Yes, that could also damage the complexes. It is usually recommended to resuspend using gentle tilting of the tubes.







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