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MTT??????????????§Ý?????3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide, MTT?????????????MTT????????????????????????????????????????????????§Ö?????????????????????????????C???????? tetrazolium????????????????formazan????formazan???????????????????????????????????????????????????????? MTT???????????????formazan???????50%??N,N-?????????????20%???????????????pH 4.7????MTT???????????????????490 nm????????OD??????????????????????????DMSO????? MTT??????????????????????????????????????????????????????????????????????????????¨¢? MTT ????????? 1????????????¨²?10???§³????????????????????????????? 1000??10000??????????96??ÈÉ??????200ul. 2:???????????????????????????3??5?????????????????????????????? 3???????????3??5???????MTT?????5mg/ml??PBS ????????4§³????????????§³?????????????????????????????????????????????????????????????????150ul DMSO????10??????????????? 4??????????490nm????????????????????????????????????????????????????????????????????????????????? ??????? ??1????????????????????? ??2?????????????????§³??10??????????????????????î•?????????????????????????? ??3??????????????????§Ó?????????????????????????????öø?Øb?????????????????? MTT??????????????0-0.7????????????¦¶???????????? 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 ????????????¦Ï?; lgIC50=Xm-I(P-(3-Pm-Pn)/4) Xm:lg ?????? I:lg(??????/???????) P:??????????? Pm:??????????? Pn:??§³???????? ??????=1-?????OD?/??????OD? ????§Ö??????§³????????????????§³?????? ???? ??96???????SMMC-7721?????MTT?????????,??¨¹????1640??????,????MTT??DMSO?????????????????200ul1640,20ulMTT,150ulDMSO ??DMSO?????????????????????DMSO?????H???????§Ò???? ??????4000???????????????????20000??/ml??MTT??20ul????????§³??????????????????????????????????150ul DMSO??????????????10?????????????? |
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