Schlaepfer W W, Lee V, Wu H L
Brain Res. 1981 Dec 7;226(1-2):259-72. doi: 10.1016/0006-8993(81)91098-2.
The relationship between mammalian neurofilament triplet proteins was studied immunologically using rabbit and guinea pig antibodies to bovine neurofilament triplet proteins. Neurofilament proteins were separated by preparative electrophoresis, each protein being isolated and re-electrophoresed to enhance purification. Antisera to 68,000 (P68), 150,000 (P150) and 200,000 (P200) dalton neurofilament proteins showed greatest activity with the corresponding protein immunogen but also revealed cross-reactivity with the other two neurofilament proteins when assessed by the ELISA method. The same antigenic inoculum elicited variable cross-reactivity, more in the guinea pig than in the rabbit. Rabbit antisera to P68 was specific in that it did not cross-react with P150 or P200. Rabbit antisera to P150 and to P200 were rendered specific by absorption with P200 and P150, respectively. By electron microscopy, isolated neurofilaments became decorated with an uniform coat of antibodies when exposed to specific antisera for each of the neurofilament proteins. By indirect immunofluorescence, each antisera showed identical patterns of tissue localization, corresponding to the distribution of neurofilaments in peripheral nerve, spinal ganglia, spinal cord, cerebellum and cerebrum. Neurofilament antigens were not detected in liver, kidney, spleen, lung, bladder, intestine, aorta, heart or tongue.
利用针对牛神经丝三联体蛋白的兔和豚鼠抗体，通过免疫学方法研究了哺乳动物神经丝三联体蛋白之间的关系。通过制备性电泳分离神经丝蛋白，每种蛋白被分离并再次电泳以提高纯度。针对68,000（P68）、150,000（P150）和200,000（P200）道尔顿神经丝蛋白的抗血清与相应的蛋白免疫原表现出最大活性，但通过ELISA方法评估时也显示出与其他两种神经丝蛋白的交叉反应性。相同的抗原接种物引发的交叉反应性各不相同，豚鼠中的交叉反应性比兔中的更多。针对P68的兔抗血清具有特异性，因为它不与P150或P200发生交叉反应。针对P150和P200的兔抗血清分别通过用P200和P150吸收而变得具有特异性。通过电子显微镜观察，当分离的神经丝暴露于针对每种神经丝蛋白的特异性抗血清时，会被一层均匀的抗体覆盖。通过间接免疫荧光法，每种抗血清都显示出相同的组织定位模式，与神经丝在周围神经、脊髓神经节、脊髓、小脑和大脑中的分布相对应。在肝脏、肾脏、脾脏、肺、膀胱、肠道、主动脉、心脏或舌头中未检测到神经丝抗原。
