中国临床药理学杂志2024，Vol.40Issue(6)：844-848,5.DOI:10.13699/j.cnki.1001-6821.2024.06.013
用蛋白质印迹评价GAPDH与β-actin作为Skp2内参蛋白的研究
Evaluation of GAPDH and β-actin as internal reference proteins of Skp2 by Western blot
摘要
Abstract
Objective To determine the appropriate internal reference protein for the Western Blot(WB)assay to detect proteins related to S-phase kinase-associated protein 2(Skp2).Methods This study was carried out with several human tumor cell lines.The intracellular Skp2 protein level was down-regulated by transient transfection of shRNA plasmid,and glyceraldehyde-3-phosphate dehydrogenase(GAPDH)and β-actin were used as internal reference proteins respectively for WB assay.The relative expression levels and correlations of GAPDH,β-actin and Skp2 were evaluated.Results When the relative expression level of Skp2 changed,the relative expression levels of GAPDH in human non-small cell lung cancer cells H1299(1.32 × 106±7.86 × 104),human cervical cancer cell HeLa(6.59 × 106±1.79 × 106)and human liver cancer cell HepG2(3.13 × 106±2.87 × 105)all changed significantly(all P＜0.05).The correlation results showed that for human non-small cell lung cancer cells A549(R=0.825 8)and H1299(R=0.906 2),and there was a certain correlation between Skp2 and GAPDH expression levels(all P＜0.05).Conclusion β-actin is more suitable to be applied as reference protein in WB assay for Skp2 protein.关键词
内参蛋白/β-肌动蛋白/甘油醛-3-磷酸脱氢酶/细胞S期激酶相关蛋白2/相对表达水平/蛋白质印迹Key words
internal reference protein/β-actin/glyceraldehyde-3-phosphate dehydrogenase/S-phase kinase-associated protein 2/relative expression level/Western Blot分类
医药卫生引用本文复制引用
孙浚源,罗微微,杨金凤,贾静..用蛋白质印迹评价GAPDH与β-actin作为Skp2内参蛋白的研究[J].中国临床药理学杂志,2024,40(6):844-848,5.基金项目
国家自然科学基金资助项目(81802622) （81802622）
浙江省自然科学基金资助项目(LY18H160067) （LY18H160067）
