Phospho-specific antibodies against S285 in HTP-3 were produced by immunizing rabbits with the synthetic peptide CNPELDEIYFpSPGR (Genscript) in which the cysteine was added for conjugation to KLH (keyhole limpet hemocyanin). Polyclonal phosphoHTP-3S285 antibodies were purified and then further absorbed against htp-3(S285A) mutant worms to reduce back- ground. The specificity of the antibody was assessed by immuno- fluorescence where lack of staining was observed in htp-3(S285A) mutants but not in WT animals.
htp-3