ѧﻯѧʵʹõȥۼñȽº͵ıԼ=Գɷ֣ƻϸĤѽϸͷϸڵĿʡǿƻ-ʡ-֬ʡ֬-֮֬ӣʹʷṹϵıԣֹʽᾧѧʵлɱ
ʵӦڶͬȥۼѡΪijЩӦãµȥۼϿ [] ƪУһЩõȥۼصӦý
ȥۼһˮβ˻źһˮͷ˻ɵлͼһAһ¶£ضŨܽˮʱȥۼӻγɽˮŲλڽڲˮⲿͼһBˣˮĻϵˮһУȥۼӵľۼĿĤܽȵһҪ [] ȥۼˮijȺˮԳȣȥۼˮdz㶨ͷˮǿɱģɾص㣬ȥۼΪࣺͣӻͣͺͷͣһض¶£ԼӵγɽŨȣ֮Ϊٽ罺ŨȣCMCȥۼٽ罺Ũʱֻеڣٽ罺ŨʱԼˮķǽ档ͬγɵ¶ȳΪٽ罺¶ȣCMTˣ¶ȺŨȥۼܽԵҪһ˵֬͵ȥۼٽ罺ŨȻϸߡ
ȥۼ ʮƣSDSƣ ƣ
ȥۼ triton X-100 ʮѿգػգ tween 20 tween 80
ȥۼ CHAPS
Һ
һȥۼķࡣ
ȥۼ
ȥۼһˮһӻӵļͷ˻ɡȥۼٽ罺ŨһڷȥۼȥۼԽǿȥۼڼͷţ˲ͨӽɫȥ
[Ŵ]
ͼ 1. ȥۼһṹıAnatrace
ʮƣSDSȥۼ
SDSһַdzЧıԼʹеĵܽ⡣ƻʵķǹۼӶʹʱԣɥʧȻܡSDS1.4:1뵰ʽϣһSDSӽ϶ӣ˼ʹƷڵȵ㣬SDSҲεʵĴʹ硣DZ㷺ʹõıԾ۱ϩӾԭڡͨΪSDSʱȫѽϸƷ뾭ɴͨ19GͷӶȷDNAȫ⡣SDSʹʱԲƻάṹ˵оҪʵĻԻʵôʱʹSDSʹȥۼʱҪעһЩΪڲͬǿȵĻҺУǵԻ֮ı䣨˵ȻŨȴ0ӵ500mMʱȥۼٽ罺ŨȻ8mM0.5mM⣬SDS¶ȽϵʱᷢΪȥۼٽ罺¶ߵһֳ֣ڼδڵ»ԡSDSԿȥƷеSDS []
ƺ͵
ƺ͵Ƽʹûһͷ˻ţҲȥۼΪǵļԻŷֲڷĸ֡ǿܽϸĤ
ʮ
ʮƣSarkosyl SarcosylҲ¹ƣһӱԼԣˮ14̼¹ҲˮȻĵԭûpHҲpH仯ӰԡȻpKaֵΪ3.6´硣⣬ڿΧpH¼ıԶdzߣԼΪ3.4 10C2 2.9 10C2N/m
Sarkosylƴ¹ȺͼᷴӦͨҴؽᾧữкʹáʹõSarkosylͨSigmaõƷʽ30%ˮҺ
Sarkosylϴˮԡ¶ϴ̺ͱԼȻױƷ䷽СɫSarkosylڷ2.78-12.9% [] Sarkosylڽ洦ҵھԡͿʴSarkosylڸƾֲҩĽʳƷҵSarkosylʳƷĴװ䣬ΪʳƷĴ䡣
SarkosylõˮԡĭȶԺͶԵǿŵ㱻㷺ӦʵоDNAת¼ΪȥۼSarkosylϸڵķ봿ڵȡӡǺͼELISA
SarkosylһҪӦǴӰܽ۵סڴ˾Escherichia coliй鵰γɰ塣SarkosylԴӰȡȻ۵ĵסͨһ0.3%Sarkosyl10%SarkosylЧܽⳬ95%ĵףͨضTriton X-100CHAPSԸ߲ʵػSarkosylܽںϵ [] ڹʹغҵȱԼܽ壬ͨϡ۵ȻһȥǿȥۼܽĵۼѷSarkosylһЧܼڸߵĵŨ۵ȸ10 [] ⣬Sarkosylڽܽ⵰۵ʱۼغҸ١ʹǰ4CµںSarkosylĿȡпԴһܡSarkosylɫ̣˱ͨϡͻȥ
ȥۼ
ȥۼͲͬǵͷ˻ûмԵˮšDZΪDZȽº͵ıԼǿƻ-֬Լ֬-֮֬ӣDzƻ-ʵӣҴȥۼʹʱԡˣȥۼʹܽͷ룬ȴ˵ʵȻԼǵáڷĤӦУǴȥۼڡͷȥۼǻʯӣGLC-1GLC-2GLC-3ǽأGDNҲб [] GDNڴӽĸĤȡATPϳø []
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ͼ 2. һЩȥۼĽṹԭıG-Biosciences
Triton
Triton X-100һӱԼҪӦڴ߳ʵСгԱ(Triton X100, Triton X114, Nonidet P40, Igepal CA-630) dzƣÿ(ֱΪ9.6, 8.0, 9.0 and 9.5ƽͻPEGͷ˻ŵĴСֲ [] Triton X100Դھϩһ鱽ˮšTriton X100ٽ罺ŨȽϵͣ˲ͨȥǵ64϶ȣڴ¶£Թ۲쵽롣
ʮѿգDDMѿ
ʮѿգDDMһձԼԽԽرҪԵˮĤס֤ڴӦУDDMCHAPSNP-40ҪЧض [] DDM֬λ㡢0.17mMĸٽ罺ŨԼڽγɵˮܽͱϸĤˮȶԷdzؼ
ѿвͬȵˮ-ѿագգҲڵʵ [-]
ػ
ɫëػƣػȡڷϸĤˮסTriton X-100ܽϸڲףⲿϸĤ
Tween
Tween-20Tween-80Ǿ֬źͳϩľɽԼٽ罺ŨȺܵͣһ㶼º͵ıԼӰ쵰ʻԣܽ⵰ҲߡDzϸѽҺij֣ͨӡøʵеϴӻҺΪǿ̶ȵؼٷԽϵĿԼȥϵIJ֡
TweenȥۼһԱTween-20Tween-80IJTween-20¹ţTween-80ţͼ3ܽDzͬص㡣ЩʱdzҪڻоУΪTween-20Tween-80Ѫòͬ []
ѧʽܶȣg/mLӦ
Tween 20 ɽ洼20ϩˮɽǴ¹PEG20ˮɽǴ¹ C58H114O26 1228 1.1 ɫɫճҺ Ӧù㷺: ΪPBSTBSϴӻҺķռELISAӡǺ߷ 鶯ϸѽ⣬Ĥܼ
Tween 80 ɽ洼80ϩˮɽǴ¹PEG80ˮɽǴ¹ C64H124O26 1310 1.06-1.09 ɫճҺ ȶijЩ֦˾͵顣
Tween-20Tween-80
ȥۼߣUV)ֹȷرTriton X100Ϊȥۼһߵıˣ280nm߲¼⵰ȻIJȷ
ȥۼ
ȥۼ ͷ˻ˮԣɸһ˳ֵԡһDZȷǿҵıԼ͵3-(3-()װ)ΣΪCHAPSٽ罺Ũȣ6mMߣͨķЧȥ2-4%Ũȵĵȵ۽ͶάӾƷƱУʹáCHAPSOCHAPSͬCHAPSOһԸǿͷ˻ţʹĿԸߡˣCHAPSOϸĤܽ⡣
Һ
ҺͬʵıԼǿԴƷǹ۵ãż-żãˮãʹʷԱԡʹأȥۼʹãڶάӾڵѧоƵʵøҺй㷺ʹá⣬ʹصʱעⲻܼƷ37϶ϣᵼµʵİ []
ڷĤȥۼ
ܶ˶ĤķȽϸȤǷĤͷʵס˵ײͬһЩ⡣ҪԭĤױˮƽҺˮҺϸĤбȽϸӵ֬ʲ㼰ˮˮΪĤܽԣӦѡٽ罺ŨȽϸߵȥۼҺҲҪ㹻ſԼ㹻ȥۼȥܽƷеĤסݴ [] ÿһĤҪһģϸĤ֬ʻͼ1C-1D)ԭϣͨ¶ȺͻҺеŨȣԼȥۼԣʹĤЧܽ⡣ʱƵĤȥۼһԵױСȥۼҺ¶ȣ֮Ϊǵ㡣¶ȵӰ죬ǵ㻹ܻҺһЩصӰ죬ͺࣨ磬Triton X114ǵ23϶ȣȥۼҺ20%ĸͣǵή4϶ȣijʵȶܸӰʱԵ÷dzҪ
ȥۼѡ
õȥۼӦܹѽϸܽеĵףԺʵоûӰ졣ҪõȻĻDZԵĵʣǵڶǵ⡣ûһȥۼеʵоʹӦͬһʵ鼼ȥۼЧĺûȡʵĵʣˣͨԺʧܾ飬ҵʵȥۼ⣬ԳʹȥۼĻҪǿǣʵȥۼҺҲҪԷֹõȥۼҺˮ
ȥۼ Da Da ٽ罺Ũ (mM) 25ۼǵ ()ƽǿȥӦ
ʮƣSDS 289 18,000 7-10 62 >100 18,000 ǿ ϸѽ, Ӿ, WB, ӽ
Triton X-100 625 90,000 0.2-0.9 100-155 65 80,000 º ø߲ⶨ, IP, ܽĤ
3-((3-̴))-1-ᣨCHAPS 615 6,150 6 10 >100 6,150 º IEP, IP
һҶNP-40 680 90,000 0.059 45-50 º IEP
ʮѿգDDM 511 0.15 98 50,000 ᾧ
Tween-20 1228 0.06 76 º WB, ø, ø߲ⶨ
ػ 1229 70,000
