neutralized

Detectionofsolubleandinsoluble

A

1–40

andA

1–42peptidesbyELISA

Procedure

1、Frozencorticeswerethawed,mincedandthenhomogenizedin50mMTris-HCl

buffercontaining2%SDS,pH8.0,andamixtureofproteainhibitors.

15mg+120llysisbuffer(25PI+1mlTris-HClcontaining2%打破英语 SDSbuffer)

+2lproteinainhibitor

注：体积比为

80

倍（

10mgtissue800ulbuffer

）

2、Homogenateswerecentrifuged(100,000g

（

46000rpm,1h,4C)and

supernatantswerestoredat-70CforadditionalanalysisofsolubleA.

3、70%formicacid（60l）wasaddedtothepelletsforextractionofSDS-insoluble

A.Afterson我的野生动物朋友 icationsampleswerecentrifuged(100,000g,1h,4C)andsupernat神仙玉 ants

werestoredforanalysisofinsolubleabeta.

4、Insolubleabetawasneutra建设银行房贷利率 lizatedofformicacidwith1.0MTris-ba/0.5M

NaH

2

P0

4

,solubleabetawasneutralizatedofbufferBfor20(3l+57l)

concentrationsofinsolubleandsoluble

注：

soluble

部分至少

40

倍稀释，

insoluble

部分用

20

倍

TPS

中和。

两部分的样品150字 均需经

bufferB

再次稀释后再用

ELISAkit

检测，具体稀释比需视情

况而定

5、A40andA42wereanalyzedusingabetaELISAkits(Invitrogen).

Plateswereanalyzedspectrophotometricallyat450nmviaamicroplatereader,and

theconcentrationsofA40andA42werecalculatedusingstandardc小幸运尤克里里谱 urvesfor

A40andA42peptidebycomparingthesample’sabsorbancewiththe

hewetweight

ofcortexregionintheoriginalhomogenate,thefinalvaluesofAwere

expresdasmicrogramspergramwetweightofcortex.

The2%SDSextractsweredilutedatleast1:40sothattheassa一只牛打一个字 ycouldbeperformed

in0.05%xtractwasneutralizedbya1:20dilutioninto1MTris

phosphatebuffer,onneedtobeprepared:

1、50mMTris-HClbuffercontaining2%SDS,pH8.0

2、70%formicacid

3、1.0MTr背景的近义词 is-ba/0.5MNaH

2

P0

4

4、BufferB

0.2g/LKCl

0.2g/LKH2PO4

8.0g/LNaCl

1.150g/LNa2HPO4

5%BSA

0.03%Tween-20

1LwithultrapurewaterandadjustthepHto7.4.