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ɫŮһƬ-ְͯô-ļĸ?

rg2019-10-14 01:40:09   c57107

ҼԇЃȺ

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.f 1 ͿԌѪ( ϡጝȞ 1:8)

.96 ELISA 1 ͿѪ

.Һwϡጲ3% pˮ

.U 96׿ԭw25PBSTϴҺ(ϡɫŮһƬ-ְͯô-ļĸ?ȡ)

.ĤѪϡҺ

.Ϳ㼲ԭ (Q )KֹҺ

.ޢͿ߲ÿѪ(̼})z

.߲Ѫ ( -})_ҺƬ

.ÿѪIgG-^øYOPDƬ

Ĝʂ ( ÑԂ )

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1.Һ:5-50µlҺ0.210µlҺ50200µlҺ50300µl 12ҺҼѡ

2.Һ ()

3.ԭw:΢ɫŮһƬ-ְͯô-ļĸ?ð(U͡V;)5K,ԇɉK,춱zѪϡጺͿԭw]:ԭw؏ʹ,,ˮ_ϴփ,Ȼßoxˮˮԡ30,,á

4.ˮ , oxˮsˮ

ԇʂ(Ñ)

1.(PH9.6)   ԇ̼}_Һz1СĴ_,z҃ȷĩ100mloxˮܽ⼴ɡ4,1ƒЧ

2. ϴҺ(PBST)     ԇ 25 s PBST Һ(нYγ,Õr΢ܻ)ßoxˮsˮ1:25ϡጡ(PH,ձNaOH{7.4)

3.Һ    ȡ1Ƭ(ԇ)100mLoxˮ,ܻ,ȡ50mLҺ, ټ1Ƭ(OPD)Ƭ,ܽ,b5mL/ ƿ 10MlMƿ ), ܹ֮-2O汣档 ǰܻ ,Rrÿ1mLҺ 10µLԇ 3% pˮ(H202) ձؼpˮ!

ġE

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1.ð_Һϡጁ 1 ÿѪ(1:1000), ELISA ɫŮһƬ-ְͯô-ļĸ?ʎ , Ҝ^ҹ

2. ԭw(ԌѪ弰zѪ) 

ͬҪ,xD1-2еҼNڿԭwϲ

    D2 96 ø˰zy 20 ݘƷֈD

S11:32

S2

S3

S4

S5

S6

S7

S8

S9

S10

+1:32

-1:4

1:64

1:64

1:8

1:128

1:128

1:256

1:256

S111:32

S12

S13

S14

S15

S16

S17

S18

S19

S20

1:512

1:64

1:1024

1:128

1:2048

1:256

1:4096

ԈD1 , U Ѫϰ 50 µl/ PBST z Ѫ1:42Bmϡ 1:512ͬr,DʾϡѪ , Ȼÿ 50 µl PBST ϡጵʹÝɫŮһƬ-ְͯô-ļĸ?߲ԭ (1:4,1ݲԭ3PBST), ԭ׼100µlʎ,,4^ҹ벡ԭѪϡጶ׃1:82ϡጡ

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3.ϴҺ (1PBST) ϴ ELISA5 ,ϴˮ˦,ٌԭwĿԭwϰDELISAϵ,ÿ50µl,371Сr

4.ͬϴ5, ˦֡ѪϡҺϡጁ1Ϳ߲Ѫ (1:1000), ÿ׼50µl,,371Сr

5.ͬϴ 5 , ˦֡ lPBST ϡÿøY (1:500), ÿ׼ 50 µl ,37 1 Сr

6. ͬϴ5,ÿ׼50µlҺ (ձؼpˮ),3715犡ÿټ50µlKֹҺKֹ,492nmLxȡֵ(OD492nmֵ)

1. ԇJɘ˜

ÿKO 4 ײԭ,ԭղκѪ,ֱPBSTϡʹÝ,/ԭͺͬELISA,50µl/,ԭOD492nm ֵ1.50.5ȡ ԌտwЧr1:10241ζԃ,ԌѪ忹wЧr1:8

2.ԭƽOD492nm ֵ50%Ӌ()

ԭ4,ȥߺOD492nmֵ,Ӌʣ2׵ƽOD492nmֵ,ٳɫŮһƬ-ְͯô-ļĸ?ֵʾ50%ČOD492nmֵ

3.

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ԲԭƽOD492nm ֵ50%,zѪϡጿOD492nmֵRֵĿמԿ,С춻RֵĿמԿ,Կ׵OD492nmֵRֵrϡጶȞԓѪĿwζȡ粡ԭƽOD492nmֵ1.2,t50%0.60ijҼzѪ1:128rOD492nmֵ0.6,tԓݴzѪĿwζȞ 1:128 Rֵ̎춃ɂζ֮g ,̎164c1128֮gtwζȡgֵ1:9Oԙzy , ɂ؏ͿֻҪҼמԿ , twζж 1:128, ɿ׾Կ , tж 鿹wζȣ1:128

ELISAwЧr1:128,ж遆 1 Ϳ߿wԡ

ELISAwЧrcoPϵ:ţ:wЧr1:128,99% oЧr1:16, oЧr1:22-90֮g,50%o