Mollat P, Zhang G Y, Frobert Y, Zhang Y H, Fournier A, Grassi J, Thang M N
INSERM U. 245, Hôpital Saint-Antoine, Paris, France.
Biotechnology (N Y). 1992 Oct;10(10):1151-6. doi: 10.1038/nbt1092-1151.
We studied several monoclonal antibodies (mAbs) raised against the 100 kD Ras GTPase activating protein (p100-GAP), which was purified from human placenta. These antibodies recognized p120-GAP and p100-GAP in native and in denatured forms. The most reactive, GP15 and GP200, both recognized distinct epitopes and did not neutralize GTPase stimulatory activity. These two mAbs were selected for a two-site enzyme immunoassay, using covalent conjugates of the antibodies coupled to the tetrameric form of acetylcholinesterase as tracer. This assay was used to quantify Ras-GAP in both normal and tumor tissues and cell extracts.
我们研究了几种针对从人胎盘中纯化得到的100 kD Ras GTP酶激活蛋白（p100-GAP）产生的单克隆抗体（mAb）。这些抗体可识别天然和变性形式的p120-GAP和p100-GAP。反应性最强的GP15和GP200都识别不同的表位，且不中和GTP酶刺激活性。选择这两种单克隆抗体用于两点酶免疫测定，使用与乙酰胆碱酯酶四聚体形式偶联的抗体共价缀合物作为示踪剂。该测定法用于定量正常组织、肿瘤组织和细胞提取物中的Ras-GAP。
