1、磷酸化的DNABiotin-DNA炔基修饰的DNA巯基修饰的DNA氨基修饰的DNA,金银纳米颗粒碳纳米管、石墨烯二氧化硅纳米颗粒量子点上转换纳米颗粒,磷酸化的DNA与TiO2-堆积,如单链DNA与石墨烯,碳纳米管静电吸附,1、Biotin-DNA,2、炔基修饰的DNA及其他生物分子,3、NH2-DNA,氨基 羧基,氨基 羧基氨基 氨基,1)EDC和NHS,For best results in two-step reactions, perform the first reaction in MES buffer (or other non-amine, non-carboxylate buf
2、fer) at pH 5-6, then raise the pH to 7.2-7.5 with phosphate buffer (or other non-amine buffer) immediately before reaction to the amine-containing molecule,氨基 羧基,DCC(N,N-Dicyclohexylcarbodiimide),氨基-氨基,1)BS(PEG)n PEG Crosslinkers - Amine Reactive,2)TSAT (Tris-succinimidyl aminotriacetate),Water,Wate
3、r,NHS esters:pH 7-9,3)DSS (disuccinimidyl suberate),4)BSOCOES (bis2-(succinimidyloxycarbonyloxy)ethylsulfone),5)DSG (disuccinimidyl glutarate),6)DTSSP (3,3-Dithiobissulfosuccinimidylpropionate),Water,Water,Water,DMSO or DMF,8)BS (Bissulfosuccinimidyl suberate),9)EGS ethylene glycolbis(succinimidylsu
4、ccinate),10)Sulfo-EGS ethylene glycolbis(sulfosuccinimidylsuccinate),7)DST (disuccinimidyl tartrate),Water,Water,DMSO or DMF,DMSO or DMF,4、巯基化的DNA,1 Au-S Ag-S2 巯基 氨基3 巯基 羧基4 巯基 羟基5 巯基 巯基,巯基-氨基,1)AMAS (N-maleimidoacetoxy succinimide ester),NHS esters:pH 7-9;Maleimides:pH 6.5-7.5;Usually 7.2-7.5。Two s
5、teps reaction,DMSO or DMF,2)BMPS (N-maleimidopropyloxy succinimide ester),DMSO or DMF,3)EMCS (N-maleimidocaproyloxysuccinimide ester),DMSO or DMF,4)Sulfo-EMCS (N-maleimidocaproyloxysulfosuccinimide ester),Water,6)Sulfo-GMBS (N-maleimidobutyryloxysulfosuccinimide ester),5)GMBS (N-maleimidobutyryloxys
6、uccinimide ester),DMSO or DMF,Water,7)BMPA (N-maleimidopropionic acid),EDC,0.1mol MES pH4.7 Maleimides react with sulfhydryls (SH) at pH 6.5-7.5,8)LC-SMCC (Succinimidyl-4-N-maleimidomethylcyclohexane-1-carboxy-6-amidocaproate),DMSO or DMF,10)Sulfo-SMPB (sulfosuccinimidyl 4-p-maleimidophenylbutyrate)
7、,9)SMPB (succinimidyl 4-p-maleimidophenylbutyrate),DMSO or DMF,Water,11)Sulfo-LC-SPDP, LC-SPDP, SPDP,巯基-羧基,巯基-羟基,异氰酸脂在水溶液中很容易水解,所以最好用有机溶剂如DMSO、DMF等。如在水相中,则要使用pH8.5的碱性缓冲溶液,避免使用含羟基的缓冲液如tris缓冲液,且要注意含羟基的分子不能有氨基和巯基,因为pH7.5顺丁烯二酰亚胺失去对巯基的特异性,会同时与氨基和巯基发生反应。,巯基-巯基,pH6.5-7.5,表征:1、定性(红外表征,如交联剂的特征峰;测颗粒的在260纳米处的紫外吸收,跑电泳,电位,粒径变化),2、定量(测颗粒或上清的紫外吸收,荧光,热重力分析),谢谢大家,