CAJ | 학술논문

目的制备PHD指蛋白 10(PHF10)磷酸化多克隆抗体并用于PHF10蛋白的研究,探讨其在胃癌发生发展中的作用与机制.方法结构预测后应用固相法合成磷酸化多肽.应用磷酸化多肽免疫家兔得到多克隆抗体,并利用Dotblot方法进行此抗体特异性效价的鉴定,结果显示PHF10最低阳性检测浓度为7.8μg/L.利用PHF10磷酸化多抗对8株胃癌细胞和胃黏膜永生化细胞PHF10蛋白磷酸化水平进行检测,胃癌细胞株PH F10磷酸化水平明显高于胃黏膜永生化细胞GES-1.结果制备出的PHF10蛋白磷酸化多克隆抗体效价较高,可用于磷酸化PHF10蛋白的鉴定.结论此多克隆抗体可用于PHF10蛋白的研究.
목적 제비PHD지단백 10(PHF10)린산화다극륭항체병용우PHF10단백적연구,탐토기재위암발생발전중적작용여궤제.방법 결구예측후응용고상법합성린산화다태.응용린산화다태면역가토득도다극륭항체,병이용Dotblot방법진행차항체특이성효개적감정,결과현시PHF10최저양성검측농도위7.8μg/L.이용PHF10린산화다항대8주위암세포화위점막영생화세포PHF10단백린산화수평진행검측,위암세포주PH F10린산화수평명현고우위점막영생화세포GES-1.결과 제비출적PHF10단백린산화다극륭항체효개교고,가용우린산화PHF10단백적감정.결론 차다극륭항체가용우PHF10단백적연구.
Objective To prepare the phosphorylational polyclonal antibody to PHD finger protein 10 (PHF10) and apply it to the study on PHF10 in the tumorigenesis and development of primary gastric cancer.Methods Phosphorylated polypeptide of PHF10 was synthesized by GL Biochem Ltd ( Shanghai).Phosphorylational polyclonal antibody was obtained by immunizing rabbits with the polypeptide.The quality of the antibody was identified by Dot blotting.The minimal detectable concentration of the antibody to PHF10 was 7.8 μg/L.Results The phosphorylational polyclonal antibody to PHF10 was obtained and proved to be effective.By using the phosphorylational polyclonal antibody to PHF10,the phosphorylation status of PHF10 was detected in gastric cancer cells and immortalized gastric epithelial cells,GES-1.The results showed phosphorylation status in PHF10 in gastric cancer cells was higher than that in immortalized gastric epithelial cells,GES-1.Conclusion The phosphorylational polyclonal antibody could be applied to the research on phosphorylation status in PHF10 and phosphorylated PHF10 was expressed higher in gastric cancer cells than in GES-1 but varied in intensity.