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Title:

Western blot ????

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Title: Western blot Author: wfm Last modified by: Lenovo User Created Date: 11/22/2005 7:07:27 AM Document presentation format: – PowerPoint PPT presentation

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Title: Western blot ????


1
Western blot ????

2
??
  • ???(blotting)?????????????,?????????????????????
  • 1975?,Southern????DNA?????????(NC?)?,???DNA?RNA???
    ????DNA?????,??Southern blotting
  • ??????????,?RNA??????????,?RNA???????Northern
    blotting ,?????????????Western blotting

3
Western Blot????
  • ????????????????????????????,????????????????

4
Western Blot????
  • ??
  • ??
  • ????
  • ????
  • ????
  • ???????
  • SDS?????
  • ????

5
(No Transcript)
6
SDS?????????(SDS-PAGE)
  • ??
  • SDS ????????????,??????????????????????.
  • ? SDS ???????,??????????????????????????,?????????
    ??????.
  • ??????? SDS ??????? 11.4 (??????),
    ????SDS???,??SDS????, ?????????????????????????

7
????
  • ????(??)?N,N-??????? ??
  • SDS
  • ???Tris???
  • TEMED
  • ????
  • Tris-????????

8
???????????
????() ??????(KD)
15 12-43
10 16-68
7.5 36-94
5.0 57-212
9
-

10
??
11
??
  • ???
  • ?????????????????????
  • ?????,?0.8mA/cm2,??1.5-2h?
  • ??
  • ??????????????,??????
  • ??????,??45min????

12
?????
  • ???S??
  • ??????,????????????????
  • ?,?????
  • ????Marker

13
??
  • ????(5)
  • BSA
  • Western Blot ????(????????)

14
???????
  • ?????????????????????,??????????????????37?????4
    ????
  • ??????,?PBS?????3?,??10min?
  • ???????????,???????, 37????,4 ????
  • ??????,?PBS?????3?,??10min?

15
???????
????????(HRP) ???
??????(AP) ???????(HRP)
16
(No Transcript)
17
(No Transcript)
18
SS9??GZ0565?????????????????
  • SS9??GZ0565?????????
  • ??SS9??????,???15?????????
  • (c) ?????????,?????????????

19
Western Blot??????
20
SDS-PAGE??
  • ??????
  • ??APS?TEMED?????
  • ???????,??????,???????????
  • ????,?????????????
  • ??????????
  • ????
  • ?????

21
  • ????????
  • ?????????
  • ???????,??????????,????
  • ????????????????????,????,?????

22
????
23
??
  • 1??????????????????????
  • 2??????????????????????
  • 3????????????
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