Numerous techniques have been developed to prepare immunoliposomes based on the nucleophilic reactivity of free amine groups of proteins or peptides. One of the most popular and commonly used methods is to covalently couple free carboxylic groups to primary amines through activation of the carboxyl groups with EDC (1-ethyl-3-[3-dimethylaminopropyl] carbodiimide). EDC, which is a so-called zero-length crosslinking agent, reacts with the carboxyl to form an amine reactive intermediate (O-acylisourea). The produced O-acylisourea can be easily displaced by nucleophilic attack from primary amino groups in the reaction mixture. However, this intermediate is unstable and hydrolyzed in aqueous solutions. In order to prevent the intermediate hydrolysis, sulfo-NHS (N-hydroxysulfosuccinimide) is added to EDC to produce a significantly more stable and more soluble active intermediate (NHS ester).
Consequently, the immunoliposomes are prepared by a two-step coupling procedure: first, activating the free carboxyl group on the antibody, peptide or protein with EDC and sulfo-NHS, and then covalently conjugating antibody to the lipids through displacement of sulfo-NHS groups by amine groups of the liposomes, as depicted below. EDC/sulfo-NHS coupling reactions are highly selective and highly efficient, and the biological activity of the protein or peptide is preserved.
ImmunoFluor™-Caproylamine is a non-PEGylated product. For other carboxylic acid reactive (PEGylated and non-PEGyalated products) and also ImmunoFluor™ products suitable for other types conjugation methods see here.
ebiomall.com
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试剂盒里有详细的说明书，告诉你样品需要多少量，每个试剂需要加入多少量，和详细的实验步骤，一般买来就可以用，不用人教。
所以你问一个样需要多少量是没法回答的，测定过程是要加很多种试剂的。
谢谢谢
现在跑tricine－sds－page电泳老是跑不好，
我刚做了bradford定量，
觉得差异与空菌对照还是挺明显的，
但是这个方法只能初略的估计，
还是不能精确定量，
做HPLC我现在还没有条件，
所以我想能不能用BCA或者lowry法先进行定量啊？
这种方法如果做出来结果的话，
能不能发个sci之类的文章啊？
希望高手们指点一下！
谢谢了
LOQ-定量限LOD-检测限
至少可以说同一种蛋白定量试剂盒并不适用于所有细胞
蛋白定量试剂盒有分Bradford，BCA，Lowry等，每一种对不同的细胞都有独特的适用范围和灵敏度。所以需要多种蛋白定量试剂盒对比检测才能让结果准确
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