抗人α-synuclein多克隆抗体的制备和鉴定

制备特异性抗人α synuclein抗血清 ,用于α synuclein相关退行性疾病及病理状态的研究。用重组人α synuclein免疫BALB/C小鼠 ,Dotblot、Westernblot及免疫组化法鉴定所获抗血清。Dotblot显示该抗血清可识别高于5 0ng的重组α synuclein。Westernblot分析表明此抗血清可识别人及大鼠脑匀浆中相对分子质量为 1 .8万的蛋白质 ,与以往报道α synuclein的相对分子质量一致。免疫组化染色显示α synuclein免疫活性物质广泛分布于皮质、海马、纹状体、杏仁核和嗅球 ,主要集中于神经元核周胞质及神经末梢。提示 :所制备的抗血清可特异性识别人及大鼠脑中的α synuclein ,该抗体对α synuclein相关疾病的研究具有应用价值

Production and Characterization of an Antiserum Against Human α-synuclein

To develop an antiserum specific to human α-synuclein for the study of α-synuclein related neurodegenerative diseases and pathological conditions, BALB/C mice were immunized with recombinant human α-synuclein. The antiserum was characterized by Dot blot, Western blot and immunohistochemistry. Dot blot showed that the antiserum could detect human recombinant α-synuclein at concentration 52ng. Western blot analysis revealed that the antiserum could recognize a 18000 protein from human and rat brain homogenates, which was consistent with the reported molecular weight of the protein. Immunohistochemical staining with the antiserum showed that α-synuclein immunoreactivity was mainly distributed in cortex, hippocampus, striatum, amygdala and olfactory bulb, and was concentrated in the nerve terminals and cytoplasms near the nuclear envelopes of neurons. The antiserum can specifically recognize the α-synucleins in human and rat brains and can be used in α-synuclein related diseases.