The detail of my immunization: I run preparative SDS-PAG with 250ug of purified protein. Then I stained the gel with copper reverse stain. The band was then cut and minced with pastel and mortar (with about 2mL of PBS). The gel was injected subcutaneously into 6 location of rabbit back. Immunization was performed at 3 weeks interval. Two rabbits were being used for immunization, but both showed no result in my WB.
Is there any problem with my method? Has anyone in forum success in producing polyclonal antibody with my protocol above? If there is antibody being produced, should it be seen in 2nd boost serum?
It is very sad for not able to get antibody after taking care the rabbit for two months...it is even worst because I am unlikely to repeat it again (unless being approved by animal ethic committee again, which takes long long time...)
Any comments are welcomed...Thanks.
Edited by dcch, 01 January 2011 - 04:10 AM.