During the past five decades, various types of chemistries have been used for conjugation of molecules such as antibodies, peptides, proteins or other reactive ligands to the surface of liposomes. In general, the conjugation can be achieved through the N-terminus, the C-terminus or the available sulfur (e.g. Fab’ fraction or thiolated antibodies). Not all chemistries have the same yield and efficiency of conjugation and often reproducing biocompatible batches can be a challenge.
Copper-free click chemistry is a fairly new chemistry that has been commercialized during the past few years. More and more click chemistry-based reagents are becoming available commercially which makes the formulation development much easier for scientists. The great advantage of this chemistry is biocompatibility since no cytotoxic copper catalyst is required. By far, click chemistry is the most efficient and easiest conjugation chemistry available for coupling of antibodies and other reactive ligands to the surface of the liposomes. The conjugation chemistry is based on the reaction of the dibenzocyclooctyne (DBCO) reagent with an azide linker to form a stable triazole. DBCO moiety can be on the antibody and azide moiety can be on liposomes and vice versa. This conjugation protocol is based on the reaction of the dibenzocyclooctyne (DBCO) group of the liposomes with an azide linker on the antibody, peptide or proteins.
There are many commercialized reagents that can be used for azide modification of proteins, peptides and antibodies. To see the list of commercialized reagents for azide modification see here.
For other reactive (PEGylated and non-PEGyalated products) ImmunoFluor™ products suitable for other types conjugation methods see here.
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首先,不否认它比大多数国产的试剂盒做得好,提取的纯度、方便度和量,都还很不错。
其次,它是日本的。so我觉得实验室用用天根的,就非常好了。拒绝日货,从实验室做起。
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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SYBRGreenMix做的,但是这里建议你用EvaGreen做,灵敏度和平行性都要好于SYBRGreen,并且如果你那是ABI或者Stratagene的PCR如果用SYBRGreen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。
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