Detection of KLIP1 protein in different cell lines by immunoprecipitation and Western blot assay. (A) GFP vector (lane 1 and 3) or GFP-KLIP1 (lanes 2, 4, and 5) were transfected into 293 cells. The cell lysate was immunoprecipitated with an anti-GFP monoclonal antibody and further immunoblotted with the anti-GFP monoclonal antibody (lanes 1 and 2), anti-KLIP1-N peptide antibody (lanes 3 and 4), or preimmune serum (lane 5). Anti-KLIP1 peptide antibody detected the fusion protein GFP-KLIP1 but not GFP alone, while preimmune serum failed to detect GFP-KLIP1 protein band. (B) In Western blot assay, the anti-KLIP1-N synthetic peptide antibody, but not the preimmune serum detected a doublet of 58 and 63 kDa in all cell lines examined.