Elution of protein from SDS-Page slice for immunization

Dr E. Buxbaum EB15 at le.ac.uk
Tue Nov 26 11:17:12 EST 1996


The best results are usually obtained by thoroughly washing the gel 
slices first in fixative and then in PBS to remove the methanol and 
acetic acid. Then homogenise the gel with PBS and incomplete Freud's 
adjuvant by passing it through a fine needle several times and inject 
subcutaneously. The gel fragments release the antigen only slowly, giving 
you a good immunisation. 

Polyacrylamide is not toxic, so provided you gave the slices a good wash 
(to remove unpolymerised acrylamide), you do not harm the rabbits with 
this method, in particular as it is usually possible to make do with 
incomplete adjuvant. 

Alternatively, you can blot the gel onto nitrocellulose and homogenise 
the nitrocellulose by sonication in PBS. This works only with pure 
nitrocellulose membranes, not with reinforced nitrocellulose and not with 
PVDF. Then proceed as described above.




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