Examination of the specificity of the two polyclonal antibodies against canine NGAL by Western blotting. (A) The recombinant NGAL protein was separated by SDS-PAGE and processed for Western blot analysis. As indicated by the arrowhead, the positive control antibody (+; antibody against histidine) as well as the sera of the mice (lane 2) and the rabbit (lane 4) immunized with NGAL were able to recognize the recombinant canine NGAL protein (~42 kDa), but the pre-immunized sera of mouse (lane 1) and rabbit (lane 3) failed to detect recombinant NGAL protein. (B) Three clinic urine samples collected from one healthy dog (lane 1) and two renal failure dogs (lane 2 and 3) were used to examine the specificity of serum obtained from mice and rabbits immunized with recombinant NGAL. Both rabbit and mouse sera were able to recognize the endogenous NGAL (~25 kDa) as indicated by the arrowheads.
Lee et al. BMC Veterinary Research 2012 8:248 doi:10.1186/1746-6148-8-248