Singh R, Singh B, Malhotra S K
Biosci Rep. 1986 Jan;6(1):73-80. doi: 10.1007/BF01145181.
A monoclonal antibody (Mab J1-31) has been produced by using human brain homogenate as immunogen in mouse. Double-label immunofluorescence microscopy on cryostat sections of human, rabbit and rat brain, reveals staining of cells that are also stained with antiserum to glial fibrillary acidic protein (GFAP, a commonly used marker protein for astrocytes). However, there is no decrease in staining due to Mab J1-31 in sections incubated in antiserum to GFAP prior to incubation with the J1-31 ascites fluid. Immunoprecipitation of aqueous and detergent extracts of brain homogenate gives a single band at 30K by SDS-PAGE followed by autoradiography. Immunoelectron microscopy shows that the J1-31 antigen is associated with the cytoskeleton. Thus, the Mab J1-31 recognizes a new protein present in GFAP positive cells (astrocytes) in the brain.
以人脑匀浆作为免疫原在小鼠体内制备了一种单克隆抗体（Mab J1-31）。对人、兔和大鼠脑的冷冻切片进行双标记免疫荧光显微镜检查，结果显示，与抗胶质纤维酸性蛋白（GFAP，一种常用的星形胶质细胞标记蛋白）抗血清染色的细胞也被染色。然而，在用J1-31腹水液孵育之前，先用GFAP抗血清孵育的切片中，Mab J1-31并未导致染色减少。通过SDS-PAGE和放射自显影对脑匀浆的水性提取物和去污剂提取物进行免疫沉淀，在30K处出现一条单带。免疫电子显微镜显示J1-31抗原与细胞骨架相关。因此，Mab J1-31识别脑中GFAP阳性细胞（星形胶质细胞）中存在的一种新蛋白。
