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polyplus寶利嘉在轉染試劑上都有哪些類型？
發(fā)布時間： 2019-08-09 點擊次數(shù)： 1310次polyplus寶利嘉是陽離子化的大分子多聚乙烯亞胺(PEI)及其衍生物，其與外源基因結合形成顆粒狀，與細胞表面的蛋白多糖結合，通過細胞"內吞作用"進入細胞。PEI-復合體具有質子海綿的特性，能吸收溶酶體的H+，在復合體中形成酸性環(huán)境使核酸酶失活，保護DNA免受核酸酶的降解。進入細胞后，JetPEI復合體囊泡腫脹破裂，將DNA釋放到細胞質中。polyplus寶利嘉的基本步驟：1、JetPEI溶液2μl和質粒DNA1μg混合5-30分鐘。2、將JetPEI-DNA復合體溶液加入到細胞液中(可以在含血清的培養(yǎng)基中)。3、檢測：N/P比率(影響轉染效率的關鍵)N/P比率是對JetPEI復合體的所帶電荷的度量。它指的是DNA分子帶負電的磷酸基團與JetPEI中N殘基的摩爾比，是衡量核酸與轉染試劑比例的參數(shù)。從理論上講，JetPEI/DNA復合體的電中性為N/P=2-3，但實際操作時，N/P=5-10效果較好。polyplus寶利嘉的轉染試劑的類型：1、JetPEI轉染試劑：JetPEI是PolyPlus-transfection的主導產品，毒性低，對細胞不會造成傷害。它能把基因轉入各種粘附或懸浮的細胞系中以及原代細胞。2、Invivo-JetPEI轉染試劑：通過靜脈、大腦、腹膜以及氣管等體內途徑進行轉染，目前已在小鼠、大鼠、鴨、豚鼠和猴等動物身上取得成功。3、TaggedJetPEIs：細胞介導的轉染試劑TaggedJetPEIs是偶聯(lián)有配體的PEI衍生物，可將基因轉入特殊的細胞。A、JetPEI-Gal&invivo-JetPEI-Gal(肝細胞介導)PEI衍生物，偶聯(lián)有半乳糖。受體細胞：細胞表面表達半乳糖特異性膜血凝素的細胞，如肝細胞。B、JetPEI-Man&invivo-JetPEI-Man(樹突狀細胞和巨噬細胞)PEI衍生物，偶聯(lián)有甘露糖。受體細胞：細胞表面表達甘露糖特異性受體的細胞，如巨噬細胞和樹突狀細胞。C、JetPEI-RGD(上皮細胞和內皮細胞)PEI衍生物，偶聯(lián)有RGD肽鏈。受體細胞：細胞表面表達整合素的細胞，如上皮細胞和內皮細胞。4、FluoJetPEI：熒光轉染試劑A、JetPEI-FluoF：PEI衍生物，偶聯(lián)熒光素FluoF，通常用于胞內定位研究，適于熒光顯微鏡和流式細胞儀的檢測。B、JetPEI-FluoR：PEI衍生物，偶聯(lián)四甲基羅丹明FluoR，常用于雙色標記和協(xié)同定位的實驗中，如用于綠色熒光蛋白或FITC標記的質粒的轉染。C、Invivo-JetPEI-FluoF&Invivo-JetPEI-FluoR：PolyPlus-transfection也提供體內轉染的熒光試劑，常用于生物分布研究。5、JetPEI-Biotin和invivo-JetPEI-Biotin：PEI衍生物，偶聯(lián)Biotin，通過鏈親和素檢測JetPEI/DNA在細胞內的定位和分布。
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