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350784478

木虫 (正式写手)

[交流] MTT原理、步骤、结果及分析

MTT??????????????л?????3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide, MTT?????????????MTT????????????????????????????????????????????????е?????????????????????????????C???????? tetrazolium????????????????formazan????formazan???????????????????????????????????????????????????????? MTT???????????????formazan???????50%??N,N-?????????????20%???????????????pH 4.7????MTT???????????????????490 nm????????OD??????????????????????????DMSO?????
MTT??????????????????????????????????????????????????????????????????????????????á?

MTT

?????????
1????????????ú?10???С?????????????????????????????
1000??10000??????????96??壬??????200ul.

2:???????????????????????????3??5??????????????????????????????

3???????????3??5???????MTT?????5mg/ml??PBS ??)20ul.
????????4С????????????С?????????????????????????????????????????????????????????????????150ul DMSO????10???????????????

4??????????490nm?????????????????????????????????????????????????????????????????????????????????

???????
??1?????????????????????
??2?????????????????С??10??????????????????????顣??????????????????????????
??3??????????????????в?????????????????????????????鲽?豣??????????????????

MTT??????????????0-0.7????????????Χ????????????

IC50?????????????????????50???????????????????????????????????????????????????????????????????????????????????????50??????????????????????IC50???????2??????????????????????????

???????????????0.1??0.01??0.001??0.0001??0.00001??0.000001?????????10?????????0.1?????????0.95?? 0.80??0.65??0.43??0.21??0.06??????
???????
Pm=0.95
Pn=0.06
P=0.95+0.80+0.65+0.43+0.21+0.06=3.1
Xm=lg0.1=-1
lgI=lg0.1/0.01=1
lgIC50=-1-1*(3.1-(3-0.95-0.06)/4)=-3.6025
IC50=0.00025

????????????ο?;
lgIC50=Xm-I(P-(3-Pm-Pn)/4)
Xm:lg ??????
I:lg(??????/???????)
P:???????????
Pm:???????????
Pn:??С????????

??????=1-?????OD?/??????OD?
????е??????С????????????????С??????

????
??96???????SMMC-7721?????MTT?????????,??ü????1640??????,????MTT??DMSO?????????????????200ul1640,20ulMTT,150ulDMSO ??DMSO?????????????????????DMSO?????H???????б????

??????4000???????????????????20000??/ml??MTT??20ul????????С??????????????????????????????????150ul DMSO??????????????10??????????????
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朝为田舍郎,暮登天子堂。将相本无种,男儿当自强。
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linzhuo

木虫 (著名写手)


好,顶一个,只是结果分析写的少了点
3楼2008-10-22 21:38:08
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mr_songhua

谢谢,可惜不能加分
2楼2008-10-22 16:20:24
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