肌肉生成抑制因子多克隆抗体的制备与鉴定

Preparation and identification of polyclonal antiserum against myostatin

运用纯化的肌肉生成抑制因子(myostatin,简称MSTN)蛋白为免疫原制备弗氏佐剂疫苗,主动免疫健壮的雄性青年家兔以制备抗血清,运用饱和硫酸胺沉淀法对所制备的抗血清成功地进行粗提.Western Blot和ELISA检测结果是:当抗原质量浓度为20μg/mL,抗体稀释800倍时,所测的P/N仍大于2,证明抗体的制备是成功的,抗体效价为1∶12800,且特异性较高.抗体稀释400倍时,血清抗体D450nm值为0.624,P/N值为5.552.

Oil-emulsion vaccines were prepared using purified MSTN protein, by which the rabbit was immunized four times to obtain MSTN-specifc antibody, which were then purified with ammonium sulfate salting-out procedure. Western-Blot and ELISA assay results showed that when the contentration of antigen was 20 μg/mL and the dilution factor was 800, the mensured P/N was larger than 2, indicating that preparation of MSTN-specific antibody was successful and the titer of antibody was 1∶(12 800). The specificity of the antibody was very high. When the dilution factor of antibody was 400, D_(450 nm) was 0.624,P/N was 5.552.