
Liposomes are extensively used to study the interaction of proteins, peptides and other molecules with the surface of a lipid membrane. One of the parameters that affects this interaction is the charge of the liposomal membrane. Liposomes are always made in aqueous environment and they are sized to the desired size in liquid state using various methods such as high-pressure extrusion through nano sized pore track etch membranes. Liposome without water is meaningless. In rare occasions, liposomes are freeze dried and proliposomes are formed in the presence of a lyoprotectant such as trehalose. Using a lyoprotectant is necessary in order to maintain the size of the liposomes after rehydration.
Phosphatidylserine (PS) is a negatively charged molecule. The amount of the negative zeta potential in the liposomes depends on the molar percentage of the phosphatidylserine (PS) in the formulation. Anionic Lyophosome™-Phosphatidylserine (PS) catalog contains products made from 0.5 up to 90 percent DOPS with various amount of negative zeta potentials. The matrix lipid, DOPC, that is used in the formulations does not significantly contribute to the charge of the liposomes. Anionic liposomes are used in blood complement studies and various types of in vitro studies.
Lyophosome™ product catalog is composed of a large selection of freeze-dried liposomes with various types of lipids and wide range of zeta potentials and different properties. Lyophosome™ products should be used by scientists who understand liposome formulation and have the proper equipment to check the size, separate non-encapsulated drugs and do the proper assays. Freeze-dried liposomes cannot be used blindly.
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可以用移液器将细胞吸出来并高速离心,沉淀重悬于PBS中洗涤,接着就可以裂解提取蛋白了。可以用超声,酶解等等,裂解后离心收集上清。
做的比较好的,一般都是上海地区的,你可以看下基尔顿生物。
我刚开始做转染,悬浮细胞,分别做过表达和敲减,看了很多文献,大都没有提及转染后是用转染的这同一批细胞同时做pcr,wb,cck8,凋亡,细胞周期;还是说这次转染只做pcr或wb,再转染一次做cck8或细胞周期。剩下的功能试验均同前,转染一次做一次?我养的是悬浮细胞,转染后做cck8这些功能试验前需要离心换液吗?跪谢解答!
细胞状态与密度;转染试剂的类型、用量;DNA的品质、用量;转染复合物的品质、作用细胞的时间长度。