iposome formulations can be broadly divided into cationic, anionic and neutral subtypes. Cellsome® liposomes made from neutral PEGylated lipids provide certain benefits. One of the most well-known Neutral PEGylated Lipids used if for the drug Doxil®. Liposomal doxorubicin (Doxil®) is a liposome formulation encapsulating the chemotherapeutic drug, doxorubicin, thereby decreasing its toxicity.
To improve liposome stability and enhance their circulation times in the blood, a sterically-stabilized, hydrophilic polymer, polyethylene glycol (PEG), has been shown to be the optimal choice for obtaining sterically-stabilized liposomes. Using a Neutral Liposome with PEG will protect the liposomes from circulating proteins, improving their plasma clearance and enhancing their therapeutic effects. Doxil® also contains polyethylene glycol (PEG)-lipid resulting in pegylation of the liposome surface, which enables it to have a prolonged circulation and a reduced volume of distribution, thereby improving tumor uptake through the enhanced permeability and retention effect (EPR) and extending effective tumor therapy.
The three products provided that are made from Neutral PEGylated Lipids provided here consist of either HSPC:Chol:DSPE-PEG(2000) (65:30:5 molar ratio), DSPC:Chol:DSPE-PEG(2000) (65:30:5 molar ratio), or DOPC:Chol:DOPE-PEG(2000) (65:30:5 molar ratio).
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用Mabteck公司的ALP标记抗体吧，配合Millipore公司的Multiscreen-IP96孔板，所有的试剂都可以减半，照样可以出非常好的结果，4*96孔的试剂盒我一直做8*96孔，但BCIP/NBT显色液没试过减半，因为它很便宜。记得当年第一次做实验就出了很好的结果，经过不断摸索，做出的蓝紫色斑点确实非常漂亮，几乎没有非特异斑点，嗯，ELISPOT是我最拿手的实验之一。
关于PBMC:
拿到血液标本后，首先我要做的是尽快分离PBMC（一般2h内），国产的分离液里天津灏洋的比较好，然后用冻存液冻存PBMC。PBMC的冻存过程比一般的细胞株要更为严格，最好用冻存器。ELISPOT试验的第一天，在包被抗体的同时，将细胞复苏，完全培养液培养过夜，第二天收集细胞并计数即可。
尤其是获得SCI文献引用的产品，
绝对ok的
请问一般的实验室可以进行ELIspot吗？（我可以买到现成的试剂盒，资金这个应该没有问题。）ELIspot的难点在哪里？多谢大虾们的指点！这个对我很重要，多谢！！！
