z*******a
发帖数: 175 | 1 I have been trying to show protein interaction between a kinase and its
predicted substrate by regular co-
immunoprecipitation. but failed twice already.
boss suggested crosslink co-IP again and again, very nicely though. I heard
it is almost impossible to publish
protein interaction data coming out from crosslink co-IP.
every Daxia,
Anybody see any published paper having crosslink co-IP data in it? If you do
, please forward me the basic info
( journal and author or title). Thanks a lot.
any ot |
W****C
发帖数: 1937 | 2 SCIENCE 2008 320(5882): 1496-1501
YASEMIN SANCAK et al.
The rag gtpases bind raptor and .....
浓度，时间（高了会使蛋白沉淀很多， 而且假阳性）要优化， 多抗比一抗好。
crosslinking很可能会大大降低单抗的亲和力 |
z*******a
发帖数: 175 | 3 Thank you so much, WPBIOC (唐僧) |
W****C
发帖数: 1937 | 4 thermo scientific有各种crossliker卖。 选的时候要考虑溶解性（水溶/脂溶），
membrane permeable, reversible or not |
z*******a
发帖数: 175 | 5 informative. Thanks a lot |
h****n
发帖数: 2552 | 6 Have you ever sonicated your cell lysis before CO-IP? |
z*******a
发帖数: 175 | |
h****n
发帖数: 2552 | 8 I suggest you not doing a sonication since it may kill some proteins. If
your cell lysis is very sticky just vigorously pipette your samples up and
down several times untill foaming.
【在 z*******a 的大作中提到】
: yes, I have.
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