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 the 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 of the linker lipid incorporated in the liposomes with EDC and sulfo-NHS, and then covalently conjugating the antibodies to the lipids through displacement of sulfo-NHS groups by antibody amines, 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™-Glutaryl is a non-PEGylated product. For other amine reactive (PEGylated and non-PEGyalated products) and also ImmunoFluor™ products suitable for other types conjugation methods see here.
ebiomall.com
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蛋白的类型为PDGF。其有一种二聚体为PDGF-BB。
请教第一点:我看了Elisa的说明书,现在检测PDGF有两种试剂盒,一种是检测PDGF的,要求的试剂样本为组织匀浆;一种是检测PDGF-BB的,要求的待测样本为血液或血浆。
我现在的实验目的是想更精确,检验组织中的PDGF-BB,但是手中的是人体韧带组织样本,非PDGF-BB试剂盒所要求的组织样本为血浆或血清,我不知道是否可以用PDGF-BB的ELISA试剂盒的来做,所以特地请教:)
请教第二点:市面上有RB公司的96T,2500元;另外有Uscnlife的Elisa试剂盒,96T的3680元,都说是原装进口的,我该如何选择呢?为何差价这么多?谢谢指点!
上海好多的供应商的,具体还是看你选择了,其实试剂盒产品都差不多,什么远慕生物,古朵生物...好多的额