【摘要】 本实验应用 Triton X-100处理,羟基磷灰石初步纯化和 DEAE 纤维素层析的方法,从牛脊髓中提取神经细丝蛋白,免疫小鼠,经细胞融合与鉴定,证明所能分泌抗神经细丝蛋白单克隆抗体杂交瘤细胞株特异性高,性质稳定,可以用于细胞学、组织学、病理学等方面的研究。更多还原

【Abstract】 Neurofilament（NF）triplet proteins were isolatedfrom bovine spinal cord by axonefloatation andisotoniclysis.The crude extraction were partiallypurified with hydroxyapatite gel by a batchwisemethod.Followed by DEAE-cellulose columnchromatography,the three components of NFproteins（NF-L,NF-M,NF-H）can be completelyseperated.The three bands of NF were immuno-stained on the transfered nitrocellulose paper.Two hybridoma clones secreting monoclonalantibodies against bovine neurofilament tripletproteins were prepared by fusion of spleen cellsfrom the mice immuninzed with crude andpurified bovine NF proteins with X63-Ag 8.653mouce myeloma cells,One hybridoma clone wasprepared by multi-well plate culture and anotherby soft agarose culture,They stained the neuronsand neuronal processces of human brain andspinal cord in frozen and paraffin embededsectinons.There two McAbs did not crossreactwith other intermidate filaments.The immunoglobulintypes of both McAbs were IgG2a（E₁）and IgM（D₂）.更多还原