Immunoprecipitation's theory

Nikolai Chitaev nikolaic at VISAR.WUSTL.EDU
Mon Jan 1 16:18:35 EST 1996


> To: methods at net.bio.net
> From: Giorgio Spagnol <spagnol at galactica.it>
> Subject: Immunoprecipitation's theory
> Date: 1 Jan 1996 19:47:20 GMT
> 
> Dear fellow researchers,
> I would be bery grateful if anyone could point to me an article
> illustrating the PRINCIPLES of the immuneprecipitation reaction. It was
> to date impossible to me NOT to find a protocol (there are at least one
> for any immunology lab manual, and in the same book, without any reasons
> the protocols may vary in two different chapters) but to find explained
> WHY you should use a particular buffer,WHAT the advantages or
> disadvantages of using protein A coupled to agarose instead of Ab linked
> to sepharose as precipitin etc, etc, etc.
> Thank you very much for any help.
> Giorgio Spagnol
> 
> 
The Principles? Nothing special - Ag/Ab interactions.
How many labs - so many protocols.
Buffer - depends where your Ag located within the cell.
Using Pr.A - disadvantage since not all Ab have high affinity toward Pr.A or
Pr.G, - this could be overcame using secondary Ab, and you clear yourself from
making a lot of trouble in attemt to prepare high quality Ab linked directly 
to solid support (whatever you choose).
As alternative, I think, anti-Fab, or anti-(Fab)2 antibodies could be used (if
you have the good source of it). Then you could either directly link them to
whatever you choose, or load them on Pr.A/G.

Basically, everything depend of what do you whant from your RIP (reaction of 
immunoprecipitation).

Nikolai
http://128.252.119.253






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