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

1)抗蛋白酶Nexin-1(PN1)多肽抗体的制备合成了14个多聚体，与预测的大鼠PN1氨基酸高抗原性多肽区域相对应。2)抗PN1抗体的特异性用从毛乳头细胞条件培养液中提纯的GST-PN1融合蛋白和天然PN1蛋白经Western印迹分析未见反应。3)PN1蛋白在毛囊中的定位用抗PN1抗体进行免疫组织化学染色，未检测到PN1蛋白在人和小鼠毛囊中的定位。抗PN1多克隆抗体由实验室赠送，用pET系统表达大鼠PN1融合蛋白。但同样的免疫组织化学检查没有检测到任何反应。这些结果表明，毛囊中PN1蛋白的数量比估计的要少得多。

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.