????????,????????,???????????(Isoelectric point pI),?????????,???????? - PowerPoint PPT Presentation

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Title:

????????,????????,???????????(Isoelectric point pI),?????????,????????

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Title: 5 Author: kayxiong Last modified by: kayxiong Created Date: 8/29/2004 1:27:56 PM Document presentation format – PowerPoint PPT presentation

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Title: ????????,????????,???????????(Isoelectric point pI),?????????,????????


1
?5? ????????
2
??????
  • ??????
  • ????????
  • ????????
  • ????????
  • ????????
  • ?????????

3
????????
  • ?????????????
  • ????????
  • ????????
  • ??????
  • ????????

4
????????
  • ????????,????????,???????????(Isoelectric point
    pI),?????????,????????
  • ????pH???,?????????????????????,?????????,????????
    ???????????,?????????,?????????????????????(Elect
    rophoresis)?

1 ?????????????
5
??
  • ???????pH???,??????????????????,?????????????

6
  • ???????????,?????????????????????????????,????????
    ?????
  • ??????????????????,??????????????????????

2 ????????
7
  • ?????????????????????????????????
  • ???????,???????????
  • ???????,??????????????

3 ????????
8
3 ????????
????
  • ??????,???????pH?????,???????????????
  • ??????,????????????,???????,??????????,???????????
    ????
  • ??????????????????,?????????????????????

9
3 ????????
?????
  • ????????,????????????????,???????????????,????????
    ???????????
  • ?????????????????????,??????????
  • ??????????????????????????????????

10
  • ??????????????????????????,????????????,??????????
    ????????????????????????(denaturation)?

4 ??????
11
??????
  • ???????????????,?????????,????????????????????,???
    ?????????????????????????????????????????????

12
  • ???????????????????????????
  • ????????280nm ??????????,???????280nm ?????????
  • ??????,?????????????

5 ????????
13
  • ??????????
  • ??????????????,??????????????
  • (1)????????????????????
  • (2)??????????????????
  • (3)????????????????????

14
  • 1.????????
  • (1)??????????????????????????????????
  • (2)????????,????????????
  • ??????????????,????????????,???????????????
  • (3)?? ?????????,?????????????????,?????????
  • (4)?????????????????
  • (5)???????????????????

15
(1)???
  • 2.????????
  • ????????????,?????????,??????????(Precipitation)?
  • ??????????????????????????????????,???????????????
    ?????
  • ???????????????

16
2.????????
  • ???
  • ????
  • ???
  • ???
  • ???

17
(1)???
???
  • 2.????????
  • ??????????????,????????????????,??????
  • ??????????NH4gtKgtNa
  • ??????????PO43-gtSO42-gtCl-
  • ?????????????????????????????,??????????????,?????
    ???????????

18
(1)???
????????
  • 2.????????
  • ???????,???????????????,?????????????,???????????,
    ??????????????
  • ???????????????????????????????????,??????????(30
    -50),?????????????

19
(1)???
???????
  • 2.????????
  • ??????????,?????,????????????,???????????
  • ??????????pH ????????????,????????????????????????
    ????pH ????????????
  • ?????????pH???????

20
(2)????
? ???
  • 2.????????
  • Dialysis?????????????????,????????????,???????????
    ?????????????????
  • ??????????????????,???????????

21
(2)????
? ???
  • 2.????????
  • ??????????????????????????,??????????(??????????)?
    ?????
  • ?????????????????

22
(2)????
?????
  • 2.????????
  • ??????????????,?????????????????????
  • ?????????????????????????
  • ??????????100??50??30??10??5??1??5??1?

23
?????
  • 2.????????

24
(3)???
???????
  • 2.????????
  • ??????????????????????????,???????????????????????
    ?????????
  • ??????????
  • ????-???
  • ????-???

25
(3)???
????????
  • 2.????????
  • ??????????????????,????????????????????????????
  • ??????????????,?????????????,???????????????????

26
(3)???
?????????
  • 2.????????
  • ?????????,?78.3Mpa???,CO2??????,???31.1C,????????
    ???,CO2???-????,??????,???????????????????????????
    ???????????

27
(3)???
?????????
  • 2.????????

28
(4)???
????????
  • 2.????????
  • ???????????????????,???????????????????
  • ?????????????????(????????),???????(????????????)?
  • ????????????????,???????????????????????????????,?
    NaCl????????,??Na???????,???????????????????

29
(4)???
????????
  • 2.????????

30
(4)???
??????
  • 2.????????
  • ??????????????????????????????????????????????????
    ?????????
  • Sephadex-?????G10-G200
  • Sepharose-?????2B,4B,6B
  • Sephacryl-?????????S100,S200,S300,S400

31
(4)???
??????
  • 2.????????

32
(4)???
?????
  • 2.????????
  • ??????????????????????,???????????????????
  • ?????????????,?????2mol/L?3mol/LNaCl????,?????????
    ?????????????????
  • ???-Sepharose?
  • ???-Sepharose
  • ??-Sepharose

33
?????
(4)???
  • 2.????????
  • ??????????????????????????????????????????????????
    ?????
  • ???????????????????????,??????????????????????????
    ???????????,??????????????????,???????????????????
    ?????,???????????????????????????,??????????????

34
?????
(4)???
  • 2.????????
  • ???????????????????????????????????????????

35
(5) ???
  • 2.????????
  • ????????,??????????????????,???????????????pH?????
    ???,???????????????????????????,???????,??????????
    ??????
  • ????????,?????????????????(v)???????(E),??????(q)?
    ?????????(f)?

36
(5) ???
?SDS-??????????
  • 2.????????
  • SDS(???????)????????????,????????????????????1 ?
    2?????
  • ??,?????????????????SDS?????????,?????????????????
    ???????????????????,??????????????,??,E?q?????????
    ?????????????????????????
  • ??????????????,????????????????

37
(5) ???
?????
  • 2.????????
  • ??????(pH3-9,???????pH5-7)???????????????????,????
    ,?????????????????,?????????????????????,????????

38
3.????????
  • ?????????
  • ????????0.2-1.7mg/L
  • ?Folin-Lowry?????????????????????????????,???????
    ,???????????,????????????????,?650nm?????????????2
    5?g-250?g/ml.

39
???????(Bradford?)?????G250???????????,??????,???
?????????,???????,?595nm??????,????????????,5-50
?g/ml.
  • ??????
  • ?????260nm?????,???????,???280nm?260nm????
  • ?????mg/ml1.45A280-0.74A260

40
4.?????????
  • ?????????????(??????????,???????)????????????????
    ????????
  • ?????????????????
  • ?????????SDS??????????????
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