溶液中NP-40对TCA沉淀蛋白影响# Biology - 生物学
J*n
1 楼
用TCA沉淀蛋白,溶解蛋白的溶液中有大概1%的去垢剂NP-40,所以在加入Na-
deoxycholate (DOC)就有很多沉淀出来,不知道会不会对蛋白得率有影响?
many thanks in advance!
这个是protocol
1. to one volume protein, add 1/100 volume of 2% DOC, keep on ice for 15 min
2. add 1/10 volume 100% TCA and keep on ice for 1 hr
3. centrifuge at 10,000 g for 10 min at 4C
4. wash the pellet in acetone to get rid of residual TCA
5. resolubilise the pellet in appropriate buffer, and add Laemmli sample
treatment buffer to protein solution
6. SDS-PAGE
deoxycholate (DOC)就有很多沉淀出来,不知道会不会对蛋白得率有影响?
many thanks in advance!
这个是protocol
1. to one volume protein, add 1/100 volume of 2% DOC, keep on ice for 15 min
2. add 1/10 volume 100% TCA and keep on ice for 1 hr
3. centrifuge at 10,000 g for 10 min at 4C
4. wash the pellet in acetone to get rid of residual TCA
5. resolubilise the pellet in appropriate buffer, and add Laemmli sample
treatment buffer to protein solution
6. SDS-PAGE