一、原代肝細(xì)胞轉(zhuǎn)染siRNA簡述
小干擾RNA轉(zhuǎn)染，將原代肝細(xì)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(shí)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(zhuǎn)染試劑轉(zhuǎn)染原代肝細(xì)胞（貨號：AD600050，Zeta life，United
States）。 以下CHOP siRNA序列用于轉(zhuǎn)5'-CAGUAUCUUGAGUCUAAUATT-3'和 5'-UAUUAGACUCAAGAUACUGTT-3' , 轉(zhuǎn)染 24 h 后，將肝細(xì)胞放置于300 μM MCT 36 h，然后對細(xì)胞活力、Annexin-V/PI 染色或蛋白質(zhì)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(xì)胞培養(yǎng)，并完成后期培養(yǎng)實(shí)驗(yàn)。
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(xué)舊金山校區(qū)聯(lián)合開發(fā)用于哺乳動(dòng)物細(xì)胞、活體動(dòng)物轉(zhuǎn)染的 Advanced DNA RNA 第三代多肽小分子轉(zhuǎn)染試劑，此技術(shù)成為新的蛋白功能、免疫細(xì)胞及干細(xì)胞治療、研發(fā)及生產(chǎn)的主要關(guān)鍵技術(shù)之一。
四、產(chǎn)品信息
