Anti-EGL-5 antibodies were raised in rabbits by injection of a ST:EGL-5 fusion protein isolated from Escherichia coli (HRP, Inc., Denver, PA). For generation of the fusion protein, the coding sequence of a full-length egl-5 cDNA was used. egl-5
A total of 700μg of purified GST::Ce-lamin protein was provided to Cocalico Biologicals (Reamstown, PA) for antibody production in two guinea pigs, CUMC-GP 27 and CUMC-GP 28. A total of four boosts were given to each guinea pig before exsanguination. lmn-1
sacy-1 cDNA sequences were cloned into the E. coli expression vector pMal-c2 to create an inducible fusion protein wherein maltose binding protein was fused to amino acids 411-578 of SACY-1 (MBP::SACY-1(411-578)). MBP::SACY-1(411-578) was column and gel-purified and used to immunize rabbits. Immunizations and sera collection were performed using standard protocols (Cocalico Biologicals, Inc., Reamstown, PA). Rabbit antibody (R217) was affinity purified and was suitable in western blots with partially purified SACY-1 preparations. sacy-1