University of Bologna
Question
Asked 11th Feb, 2021
Does sds interfere with immunization?
I want to extract a protein from polyacrylamide gel to be used for immunization. I am wondering if I need to run PAGE since sds might interfere with the immunization process.
Thank you
Most recent answer
I assume this means no interference from SDS. I intend to use reverse staining method and elute the protein from gel. Thank you
All Answers (4)
Private Person
The classical procedure is to run your sample on SDS-PAGE, fix, stain and destain the gel and then cut out the band of interest. Wash with 2 changes of PBS, mash it up thoroughly in adjuvants (complete first time, incomplete for boosters) and then inject. Contrary to acrylamide, polyacrylamide is not toxic, but serves as a depot for the antigen. If you are very short of antigen, you can inject directly into a lymph node at the hind leg of the rabbit, this is a surgical procedure, so you'll need a vet to carry it out.
For all antibody-related procedures, check out ISBN 9780879693145, which is still the authoritative reference. And of course, you need permission from your local animal use committee for any such experiments.
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University of Bologna
Masood Sepehrimanesh
Thank you for your comment, Masood. I'm aware of other purification methods, but I have concluded this suits me the best.
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