兔抗人NESTIN抗血清的制备与鉴定(简报)

巢蛋白(nestin)属Ⅵ类中等纤维蛋白。最初在神经系统发育早期发现有该蛋白的表达。后在正常神经系统中发现仅在未分化的神经前体细胞中有nestin的短暂表达,分化后在神经元和神经胶质细胞中分别被NF(neurofilament,神经丝)和GFAP(Glial fibrillary acidic protein,神经胶质元纤维酸性蛋白)所代替。所以nestin的表达通常被视为神经前体细胞的标志之一。就我们所知,目前商品化nestin的抗体都是鼠抗鼠(克隆Rat401)的抗体,而人鼠之间该蛋白序列的同源性仅为50%左     

兔抗人NESTIN抗血清的制备与鉴定（简报）

Nestin belongs to the class VI intermediate filament family and it is a marker for neural progenitor cells. In this work, the 3'-terminal coding sequence(396 bp) of human nestin gene was cloned into pGEX-3X plasmid and introduced into BL21 E. coli cells. The GST-nestin protein was purified with an affinity column. Anti-human nestin antiserum was raised by immunizing a rabbit with the fusion protein. The high specificity of the antibody against human nestin was confirmed by western-blot and immunocytochemistry analysis.