摘要
成年大鼠瓦勒氏变性神经来源的雪旺氏细胞（ＳＣ），经体外无血清分离培养，收集ＳＣ无血清条件培养液，用ＳＤＳ－ＰＡＧＥ分析其超滤浓缩液（分子量大于１０ＫＤａ）中ＳＣ分泌蛋白，发现其含有１４条蛋白带，分子量从大于９４ＫＤａ至小于３４ＫＤａ。用免疫印迹技术、抗２．５Ｓ神经生长因子（ＮＧＦ）抗体鉴定证实ＳＣ分泌蛋白中含有ＮＧＦ。用Ｄｉｓｃ－ＰＡＧＥ（４℃）分离ＳＣ分泌蛋白带，分别经电洗脱、浓缩收集１０份蛋白区带，结合有髓腹侧运动神经元细胞培养、ＮＴＦｓ生物学活性鉴定，初步发现其中一组蛋白区带（Ｂ＋）含运动性神经营养因子（ＮＴＦ）活性，其分子量在６５ＫＤａ至３４ＫＤａ。
Abstract The dissociated Schwann cells(SCs), taken from the adult rat sciatic nerve undergoing Wallerian degenration, were cultured in serum-free medium. The SCs serum-free conditioned medium (SC-SF-CM) was collected, ultrafiltrated and concentrated. The proteins secreted by SCs in SC-SF-CM ultrafiltrated-concentrated solution were analysed by SDS-PAGE.It was found that there were 14 protein bands, which molecular weight. were from more than 94KDa to less than 43 KDa, and main band of them was at 65KDa. By immunoblotting (Western Blot) for anti2.5S Nerve Growth Factor (NGF) antibody, NGF was identified to exist in the porteins secreted by SCs. SC-SF-CM ultrafiltrated-concentrated solution was also separated by Disc-PAGE at 4℃,then electroluted, concentrated and colleted. By biological activity exa mination for Motor Neurotrophic Substances, one protein zone (B+) of them was identified to have motor NTF activity, which contained five protein bands from 65KDa to 34KDa.It is necessory to further purify.
出处
《中华显微外科杂志》
CSCD
北大核心
1994年第3期201-203,共3页
Chinese Journal of Microsurgery
基金
国家教委博士点基金
