Zeta Life Advanced转染试剂高效率转染大鼠原代肝细胞(IF6.684)

2023-04-21

一、原代肝细胞转染siRNA简述

小干扰RNA转染,将原代肝细胞接种到6孔板或96孔板的含10% FBS Eagle培养基中,培养二十四几小时后,用100 nM的CHOP siRNA 或100 nM 阴性对照 siRNA (siNC) 使用美国Zeta Life公司Advanced DNA RNA转染试剂转染原代肝细胞(货号:AD600050,Zeta life,United


States)。 以下CHOP siRNA序列用于转5'-CAGUAUCUUGAGUCUAAUATT-3'和 5'-UAUUAGACUCAAGAUACUGTT-3' , 转染 24 h 后,将肝细胞放置于300 μM MCT 36 h,然后对细胞活力、Annexin-V/PI 染色或蛋白质印迹测定。

培养条件:选择美国Zeta Life公司胎牛血清(#z7181FBS-500)对原代肝细胞培养,并完成后期培养实验。


二、Small Interfering RNA Transfection 原文分享

Hepatocytes were seeded into 6-well plates or 96-well plates in Dulbecco’s Modifified Eagle Medium with 10% FBS. Twenty-four hours later, they were transfected with 100 nM of CHOP siRNA or 100 nM of negative control siRNA (siNC) using Advanced Transfection Reagent (Catalog No.: AD600050, Zeta life, United States) according to the manufacture’s protocol. The followingCHOP siRNA sequences were used for transfection: 5′-CAG

UAUCUUGAGUCUAAUATT-3′ (sense) and 5′-UAUUAG ACUCAAGAUACUGTT-3′ (antisense) After 24 h transfection, hepatocytes were exposed to 300 μM of MCT for another 36 h, followed by the cell viability assay, Annexin-V/PI staining, or western blotting


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