Fab production

[Jorg Kirberg]

In article <elc.1.0011379D at mail.nerc-oxford.ac.uk>,
elc at mail.nerc-oxford.ac.uk (Emma Louise Cleal) wrote:

> INSOLUBLE PAPAIN
> 
> Does anybody have a protocol for making Fab fragments from IgG from Ascites 
> fluid using Sigma's insoluble papain?
> 
> I'd be grateful if people could contact me if they have used this.  
> 
> Emma Cleal
> elc at mail.nerc-oxford.ac.uk
> Institute of Virology and Environmental Microbiology
>  

Ciao Emma,

I dont know why people use the papain fixed to a carrier, as it is not much
more work (and actually much more cheaper) if using papain, where one does not
need the highest quality, just itself.
I used papain from Calbiochem, #5125, and had good results; probably the 2x
cristallized powder from sigma or any other company will do as good.
The protocol I used was taken from Harlow and Lane, Antibodies. A Laboratory
Manual, Cold Spring Harbor Laboratory, 1988 and Goding, Monoclonal Antibodies:
Principles and practise, Academic press, 1986.

Good luck
jorg

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Joerg Kirberg                     EMAIL: kirberg at bii.ch
Basel Institute for Immunology    FAX:   41-61-605 13 72
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