一、原代肝細胞轉染siRNA簡述
小干擾RNA轉染，將原代肝細胞接種到6孔板或96孔板的含10% FBS Eagle培養(yǎng)基中,培養(yǎng)二十四幾小時后，用100 nM的CHOP siRNA 或100 nM 陰性對照 siRNA (siNC) 使用美國Zeta Life公司Advanced DNA RNA轉染試劑轉染原代肝細胞（貨號：AD600050，Zeta life，United
States）。 以下CHOP siRNA序列用于轉5'-CAGUAUCUUGAGUCUAAUATT-3'和 5'-UAUUAGACUCAAGAUACUGTT-3' , 轉染 24 h 后，將肝細胞放置于300 μM MCT 36 h，然后對細胞活力、Annexin-V/PI 染色或蛋白質印跡測定。
培養(yǎng)條件：選擇美國Zeta Life公司胎牛血清（＃z7181FBS-500）對原代肝細胞培養(yǎng)，并完成后期培養(yǎng)實驗。
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
三、美國Zeta Life 公司與美國加利福尼亞大學舊金山校區(qū)聯合開發(fā)用于哺乳動物細胞、活體動物轉染的 Advanced DNA RNA 第三代多肽小分子轉染試劑，此技術成為新的蛋白功能、免疫細胞及干細胞治療、研發(fā)及生產的主要關鍵技術之一。
四、產品信息
