白蜡虫脂酰辅酶A还原酶多克隆抗体的制备

Preparation of Polyclonal Antibody against Ericerus pela FAR1 Peptide

摘要: 白蜡虫脂酰辅酶A还原酶（FAR1）参与了体表蜡泌物的形成过程，为了进一步研究FAR1的功能，通过预测分析基因 far1 编码的蛋白结构，选取亲水性强、特异性好的一段合成多肽作为抗原肽，免疫新西兰大白兔制备多克隆抗体，并以BCA蛋白浓度试剂盒测定抗体浓度，通过SDS-PAGE考马斯亮蓝染色观察纯化抗体的纯度，以ELISA检测抗体效价。结果表明，纯化所得的多克隆抗体纯度高，抗体浓度0.7 mg·mL-1，抗体效价为 1：512 000，为后续FAR基因功能的研究奠定基础。

Abstract: Fatty acyl-CoA reductase(FAR1) plays a key role in the white wax secretion of Ericerus pela. In present study, the artificially synthesized polypeptide of E. pela FAR1 was used as antigen for the generation of polyclonal antibody. The obtained antibodies were purified by using activated sepharose and analyzed by SDS-PAGE. The concentration and titer of the polyclonal antibody were determined by BCA KIT and ELISA respectively. The results indicated that the titer (1:512000) and the concentration (0.7 mg·mL-1) of anti-FAR1 polyclonal antibody could meet the requirements for later experiments, and the polyclonal antibody could be used to further study the location of FAR1 protein in the later research.