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Western Blot原锟斤拷锟斤拷B
Western Blot锟斤拷WB锟斤拷锟斤拷锟斤拷锟斤拷锟斤拷锟斤拷锟斤拷印锟紼锟斤拷,锟斤拷锟角讹拷锟斤拷,锟斤拷锟斤拷锟斤拷锟絲锟統(c锟斤拷)锟斤拷锟阶憋拷锟絖(d锟斤拷)锟斤拷一锟絅锟斤拷(j锟斤拷ng)锟戒、锟斤拷效锟侥硷拷锟絞(sh锟斤拷)锟斤拷锟斤拷,锟斤拷western blot锟角革拷锟斤拷(j锟斤拷)锟斤拷原锟斤拷锟絯锟斤拷锟截拷锟皆結(ji锟斤拷)锟较檢锟統(c锟斤拷)锟斤拷(f锟斤拷)锟絪锟斤拷品锟叫碉拷某锟絅锟斤拷锟阶的凤拷锟斤拷,锟斤拷Western blot锟斤拷锟斤拷锟斤拷锟絲锟接撅拷凸锟斤拷锟斤拷锟斤拷邷y(c锟斤拷)锟斤拷锟斤拷锟絞(sh锟斤拷)锟斤拷锟紸(ch锟斤拷)锟较發(f锟斤拷)展锟斤拷锟斤拷(l锟斤拷i)锟斤拷一锟絅锟铰碉拷锟斤拷锟斤拷锟斤拷锟斤拷锟斤拷锟絞(sh锟斤拷),锟斤拷锟斤拷锟斤拷western blot锟斤拷锟斤拷SDS-PAGE锟侥高分憋拷锟斤拷锟酵癸拷锟斤拷锟斤拷锟竭測(c锟斤拷)锟斤拷锟侥革拷锟截拷锟皆猴拷锟斤拷锟斤拷锟皆ｏ拷锟紽(xi锟斤拷n)锟窖成為蛋锟阶凤拷锟斤拷锟斤拷一锟絅锟斤拷規(gu锟斤拷)锟斤拷锟絞(sh锟斤拷),锟斤拷Western blot锟斤拷锟斤拷锟斤拷锟絙锟斤拷某锟絅锟斤拷锟斤拷,锟斤拷锟斤拷锟杰岋拷(du锟斤拷)锟斤拷锟斤拷锟組(j锟斤拷n)锟叫讹拷锟皆和半定锟斤拷锟斤拷锟斤拷锟斤拷
Western Blot锟斤拷锟斤拷锟角筹拷锟斤拷锟斤拷,锟斤拷锟斤拷锟斤拷锟(sh锟斤拷)锟(y锟斤拷n)锟揭撅拷锟竭傦拷Western Blot锟侥岋拷(sh锟斤拷)锟(y锟斤拷n)?z锟斤拷i)锟斤拷锟,锟斤拷锟斤拷锟斤拷要锟矫碉拷一锟捷憋拷锟斤拷锟斤拷锟斤拷锟斤拷锟絣锟斤拷锟斤拷锟斤拷,锟斤拷锟芥實(sh锟斤拷)锟斤拷锟絕(gu锟斤拷n)锟侥岋拷(sh锟斤拷)锟(y锟斤拷n)锟結(ji锟斤拷)锟斤拷,锟斤拷锟斤拷要投锟斤拷锟斤拷锟斤拷臅r(sh锟斤拷)锟絞锟酵撅拷锟斤拷锟斤拷锟斤拷锟絑锟斤拷锟絖(d锟斤拷)锟斤拷锟斤技锟絞(sh锟斤拷)锟剿員,锟斤拷锟节橈拷品處锟斤拷,锟斤拷锟紻(zhu锟斤拷n)印锟酵匡拷锟絯稀釋憋拷锟斤拷锟饺诧拷锟斤拷锟较撅拷锟斤拷锟斤拷锟斤拷募锟斤拷桑锟斤拷軌锟?y锟斤拷n)锟酵?h锟斤拷)锟侥橈拷品锟斤拷锟斤拷锟斤拷锟狡岋拷(sh锟斤拷)锟(y锟斤拷n)锟斤拷锟斤拷,锟斤拷
Western Blot锟斤拷锟絞(sh锟斤拷)路锟斤拷(xi锟斤拷n)
Western Blot锟斤拷锟斤拷(w锟斤拷)锟斤拷(y锟斤拷u)锟斤拷(sh锟斤拷)
1. 锟竭分憋拷锟绞碉拷锟接撅拷锟斤拷g(sh锟斤拷)锟斤拷锟桔憋拷烯锟斤拷锟斤拷锟斤拷锟絲锟斤拷polyacrylamide gel,锟斤拷PAGE锟斤拷锟窖凤拷锟接篩锟斤拷锟矫猴拷電猴拷效锟斤拷(y锟斤拷ng)锟結(ji锟斤拷)锟斤拷锟斤拷同一锟斤拷锟斤拷锟叫ｏ拷锟絖(d锟斤拷)锟斤拷锟斤拷锟竭碉拷锟絗锟斤拷锟斤拷,锟斤拷
2. 锟截拷锟斤拷锟叫的匡拷原-锟斤拷锟絯锟斤拷锟斤拷(y锟斤拷ng)锟斤拷锟斤拷原锟斤拷锟絯锟侥結(ji锟斤拷)锟较岋拷(sh锟斤拷)锟絴(zh锟斤拷)锟斤拷锟角匡拷原锟斤拷位锟絚锟斤拷锟絯锟斤拷變锟絕(q锟斤拷)锟叫匡拷原锟結(ji锟斤拷)锟斤拷锟絚(di锟斤拷n)之锟絞锟侥結(ji锟斤拷)锟斤拷,锟斤拷锟斤拷锟节冿拷锟斤拷锟节伙拷锟絎(xu锟斤拷)锟結(ji锟斤拷)锟斤拷(g锟斤拷u)锟酵匡拷锟絞锟斤拷(g锟斤拷u)锟斤拷锟较呈伙拷锟絘(b锟斤拷)锟絇(gu锟斤拷n)系锟斤拷锟斤拷锟皆匡拷原锟絚锟斤拷锟絯锟侥結(ji锟斤拷)锟较撅拷锟叫高度碉拷锟截拷锟斤拷,锟斤拷
3. 1-5ng锟叫等达拷小锟侥靶碉拷锟阶ｏ拷锟缴檢锟統(c锟斤拷)锟斤拷锟斤拷锟斤拷1锟斤拷5ng锟斤拷锟斤拷涂傻锟10锟斤拷100pg锟斤拷锟叫等达拷小锟侥靶碉拷锟斤拷,锟斤拷
Western Blot锟斤拷锟斤拷(w锟斤拷)要锟斤拷
锟斤拷锟斤拷锟絩锟斤拷锟斤拷锟絖锟斤拷锟斤拷牡锟斤拷讟锟狡凤拷锟斤拷叩锟斤拷状锟斤拷锟揭猴拷锟斤拷锟斤拷锟斤拷诐锟饺诧拷锟斤拷锟斤拷 2μg/μl锟斤拷锟斤拷锟斤拷50μl锟斤拷锟斤拷锟斤拷,锟斤拷锟斤拷锟竭诧拷锟斤拷锟斤拷50mg锟組锟斤拷锟斤拷品,锟斤拷锟斤拷锟竭诧拷锟斤拷锟斤拷5×10锟斤拷6锟轿凤拷锟斤拷(g锟斤拷)锟斤拷(x锟斤拷)锟斤拷锟侥硷拷(x锟斤拷)锟斤拷锟斤拷品锟斤拷
锟(y锟斤拷ng)锟皆岋拷(du锟斤拷)锟秸橈拷品锟斤拷锟斤拷锟斤拷校锟,锟斤拷
锟絲锟統(c锟斤拷)目锟侥碉拷锟阶碉拷一锟斤拷锟斤拷锟皆傦拷锟斤拷锟斤拷锟斤拷锟角筹拷锟矫碉拷锟斤拷锟絑锟斤拷锟絖(d锟斤拷)锟斤拷(hu锟斤拷)锟叫傦拷貨,锟斤拷锟斤拷锟斤拷锟矫的匡拷锟斤拷锟斤拷锟絑锟斤拷锟絖(d锟斤拷)訂購(g锟斤拷u)锟斤拷锟斤拷
Western Blot锟斤拷锟紼
1,锟斤拷锟斤拷锟斤拷锟絴(zh锟斤拷)锟斤拷锟斤拷
A,锟斤拷锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟斤拷(du锟斤拷)锟斤拷锟組锟斤拷锟斤拷品锟斤拷取锟絤锟斤拷锟斤拷250-500mg锟斤拷锟斤拷锟絩锟組锟斤拷锟斤拷品,锟斤拷锟斤拷1ml锟斤拷锟斤拷锟斤拷酶锟斤拷锟狡勶拷锟侥匡拷锟斤拷锟阶筹拷锟斤拷試锟斤拷锟斤拷锟斤拷说锟斤拷壮锟斤拷锟皆囷拷锟斤拷锟,锟斤拷锟斤拷{锟斤拷锟斤拷峥傦拷锟斤拷住锟
B,锟斤拷锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟斤拷(du锟斤拷)锟斤拷榧(x锟斤拷)锟斤拷锟斤拷品,锟斤拷每锟捷橈拷品取1×10锟斤拷6锟轿凤拷锟斤拷1×10锟斤拷7锟轿凤拷锟斤拷(x锟斤拷)锟斤拷锟斤拷PBS锟斤拷洗锟斤拷(x锟斤拷)锟斤拷,锟斤拷去PBS锟斤拷0.1ml~1ml锟斤拷锟斤拷锟斤拷酶锟斤拷锟狡勶拷锟侥匡拷锟斤拷锟阶筹拷锟斤拷試锟斤拷锟斤拷锟斤拷说锟斤拷壮锟斤拷锟皆囷拷锟斤拷锟斤拷锟斤拷峥傦拷锟斤拷锟,锟斤拷
2,锟斤拷锟斤拷锟斤拷锟絴(zh锟斤拷)锟斤拷锟斤拷
锟斤拷KCTMBCA锟斤拷锟斤拷锟絴(zh锟斤拷)锟斤拷锟斤拷試锟斤拷锟叫诧拷锟斤拷锟絝(shu锟斤拷)锟斤拷锟斤拷锟斤拷锟斤拷锟統(c锟斤拷)锟斤拷锟斤拷品锟斤拷锟,锟斤拷
3,锟斤拷變锟皆聚憋拷烯锟斤拷锟斤拷锟斤拷锟紹锟絤(x锟斤拷)锟斤拷锟絲锟接(SDS-PAGE)
锟斤拷锟斤拷(zh锟斤拷n)锟斤拷玫臉锟狡芬猴拷锟斤拷锟斤拷锟斤拷貥锟(bi锟斤拷o)記锟侥碉拷锟斤拷锟絴(zh锟斤拷)锟斤拷锟斤拷锟斤拷锟斤拷(bi锟斤拷o)锟斤拷(zh锟斤拷n)锟街別锟较樣ｏ拷锟斤拷(bi锟斤拷o)锟斤拷(zh锟斤拷n)锟斤拷锟組(j锟斤拷n)锟斤拷一锟斤拷(g锟斤拷)锟斤拷锟斤拷,锟斤拷锟接撅拷锟斤拷x锟斤拷锟斤拷,锟斤拷
4锟斤拷锟斤拷锟斤拷锟絴(zh锟斤拷)锟紻(zhu锟斤拷n)锟狡碉拷锟斤拷锟斤拷锟絯锟絊膜锟斤拷PVDF膜
5,锟斤拷膜锟侥凤拷锟絔锟酵匡拷锟絯锟斤拷锟斤拷
A,锟斤拷膜锟斤拷5锟斤拷BSA锟斤拷液锟斤拷锟揭溫凤拷锟斤拷1小锟絩(sh锟斤拷)锟皆凤拷锟絔膜锟较的凤拷锟截拷锟結(ji锟斤拷)锟较★拷
B,锟斤拷锟斤拷锟絔锟絕(gu锟斤拷)锟斤拷膜锟斤拷锟斤拷一锟斤拷(j锟斤拷)锟斤拷锟絯锟揭溫凤拷锟斤拷1.5小锟絩(sh锟斤拷),锟斤拷锟斤拷原锟斤拷锟絯锟結(ji锟斤拷)锟较★拷
C,锟斤拷锟斤拷锟斤拷HRP锟斤拷(bi锟斤拷o)記锟侥讹拷锟斤拷(j锟斤拷)锟斤拷锟絯锟皆結(ji锟斤拷)锟斤拷一锟斤拷(j锟斤拷)锟斤拷锟絯锟斤拷HRP锟斤拷(bi锟斤拷o)記锟侥匡拷锟斤拷锟斤拷锟截匡拷锟絯锟皆結(ji锟斤拷)锟较凤拷锟斤拷锟斤拷锟斤拷(bi锟斤拷o)锟斤拷(zh锟斤拷n),锟斤拷锟揭溫凤拷锟斤拷膜1小锟絩(sh锟斤拷)锟斤拷锟斤拷锟斤拷HRP锟斤拷(bi锟斤拷o)記锟斤拷GAPDH锟斤拷锟絯锟斤拷同锟絩(sh锟斤拷)锟絲锟統(c锟斤拷)GAPDH锟斤拷锟斤拷,锟斤拷
6,锟斤拷锟結(ji锟斤拷)锟斤拷锟絲锟統(c锟斤拷)
锟斤拷锟絎(xu锟斤拷)锟絣(f锟斤拷)锟解法锟絲锟統(c锟斤拷)锟斤拷膜锟絚锟斤拷锟絎(xu锟斤拷)锟絣(f锟斤拷)锟斤拷锟斤拷锟斤拷锟斤拷,锟斤拷锟斤拷(j锟斤拷ng)X锟絲片锟截癸拷锟紷影,锟斤拷锟紻片锟斤拷锟借保锟斤拷锟斤拷锟絏锟侥硷拷锟斤拷锟斤拷锟斤拷ImageJ锟斤拷锟斤拷軟锟斤拷锟斤拷锟紻片锟斤拷每锟斤拷(g锟斤拷)锟截拷锟絣锟斤拷锟揭讹拷值锟侥旓拷(sh锟斤拷)锟街伙拷,锟斤拷
7,锟斤拷锟斤拷(sh锟斤拷)锟斤拷(j锟斤拷)锟斤拷锟斤拷
目锟侥碉拷锟阶的灰讹拷值锟斤拷锟斤拷?x锟斤拷n)?n锟斤拷i)锟斤拷GAPDF锟侥灰讹拷值锟斤拷校锟斤拷锟絗锟筋，锟斤拷锟矫結(ji锟斤拷)锟斤拷锟斤拷锟斤拷某锟斤拷品锟斤拷目锟侥碉拷锟斤拷锟斤拷锟斤拷(du锟斤拷)锟斤拷锟斤拷,锟斤拷
8,锟斤拷锟结供锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟斤拷(b锟斤拷o)锟斤拷
锟斤拷锟斤拷詳锟斤拷(x锟斤拷)锟侥岋拷(sh锟斤拷)锟(y锟斤拷n)锟斤拷锟斤拷锟斤拷Western Blot锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟結(ji锟斤拷)锟斤拷,锟斤拷
Western Blot锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟斤拷锟斤拷锟斤拷(n锟斤拷i)锟斤拷
锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟斤拷锟斤拷锟斤拷(n锟斤拷i)锟捷帮拷锟斤拷锟斤拷锟斤拷锟阶濓拷锟,锟斤拷锟絲片锟斤拷膜,锟斤拷锟皆硷拷锟揭度凤拷锟斤拷锟斤拷锟斤拷锟斤拷锟揭拷锟斤拷锟斤拷锟(sh锟斤拷)锟(y锟斤拷n)锟斤拷(b锟斤拷o)锟斤拷,锟斤拷
锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟斤拷锟斤拷锟紻片锟斤拷 
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