Effect of anti-protease nexin-1 antibody to cyclic growth of hair follicles

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1) Production of anti-protease nexin-1 (PN1) peptide antibodyFourteen mer of peptides, which is corresponding to one of the predicted peptide regions with high antigenicity of rat PN1 amino acids, were synthesized. Two rabbits were immunized with the peptides conjugated with KLH as a carrier protein, and the antiserum was prepared.2) Specificity of the anti-PN1 antibodyNo reaction was detected with Western blot analysis between the anti-PN1 antibody and GST-PN1 fusion protein or native PN1 protein, which was purified from the conditioned medium of the cultured dermal papilla cells. No reaction was detected even when using the lgG purified from the antiserum.3) Localization of PN1 protein in hair folliclesNo reaction was detected with immunohistochemical examination with the anti-PN1 antibody in order to localize PN1 protein in hair follicles of human and mouse. Anti-PN1 polyclonal antibody, for which rat PN1 fusion protein expressed by pET system, was kindly gifted from a laboratory. But no reaction was detected with the same immunohistochemical examination.These results suggested that an amount of PN1 protein in a hair follicle was much less than estimated.