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 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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请采纳。。。。
Clontech的非常好, 感觉. 效果不好可能是RNA问题, 或RTase的问题. 因为目前clontech的RTase是Takara的产品，而非原来clontech的产品，可单购。
3′-Full RACE Core Set Ver.2.0D314 20 次 1,800 元
5′-Full RACE Kit D315 RT反应10次 PCR反应50 次 3,980 元
takara的做得出来不，请各位高手指教，谢谢了！
步骤：1，RNA抽提（同RT）
2，逆转录，使用合成的oligo
3，p内参
4，使用根据已知序列设计的上游引物和提供下游引物做PCR，注意设计上游引物是退火温度与下游引物相差不大。
5，产物切胶回收，TA克隆测序
这个oligo和下游引物是我根据Takara和BD公司的manual自己设计的，经过我的使用效果很好。具体操作参照takara的3－racemanual。你先做3－race，了解熟悉下，成功后再做5－race。5－race要买试剂盒做，较为复杂！
我也想问一下。
