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.
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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。
1.模板提取(一般为RNA):Trizol、氯仿、异丙醇、无水乙醇、DEPC处理水
2.模板浓度测定:分光光度计或NanoDrop
3.逆转录:逆转录试剂盒(或者一步法试剂盒),这一步可以用普通PCR做,也可以用水域做。
4.荧光定量PCR试剂:通常有用SYBR Green Mix做的,但是这里建议你用EvaGreen做,灵敏度和平行性都要好于SYBR Green,并且如果你那是ABI或者Stratagene的PCR如果用SYBR Green还需要加一步Rox很麻烦。
5.其他:除了以上的那些还需要离心管、PCR管或板(Axygen反应比较好)、移液枪等,暂时就想到这么多。
EliKine™ 人 肝细胞生长因子 ELISA定量试剂盒
EliKine™ 人 干扰素-α ELISA定量试剂盒
EliKine™ 人 干扰素-γ ELISA定量试剂盒
EliKine™ 人 白介素-1α ELISA定量试剂盒
EliKine™ 小鼠 白介素-22 ELISA定量试剂盒
EliKine™ 小鼠 转化生长因子-β1 ELISA定量试剂盒
EliKine™ 小鼠 肿瘤坏死因子-α ELISA定量试剂盒
EliKine™ 小鼠 血管内皮生长因子 ELISA定量试剂盒
EliKine™ 大鼠 干扰素-γ ELISA定量试剂盒
EliKine™ 大鼠 白介素-1β ELISA定量试剂盒
EliKine™ 人 促甲状腺激素 ELISA定量试剂盒
只要引物设计的可以在基因组上做,那么理论上是没问题的!
只需要用RT-PCR荧光定量试剂盒里面的第二个,专门用来做Real-timePCR的那些试剂就可以了!

