a****e
发帖数: 128 | 1 想知道这种方法到底可不可靠，fresh section。我想在wt 和 mt之间 比较一种蛋白的
表达量，但貌似每次的结果都不一样，有时是wt高，有时是mt高，表达量差别又不大，
所以很难有一个确定的结论。
请问高手有没有什么建议没有，怎样才能处理这种情况? 另外，显微镜的放大倍数会对
结果又怎样的影响？目前做wester-blot confirm 不是太现实. |
q******g
发帖数: 3858 | 2 染色的时候要一起做，条件完全一样。拍照的时候，曝光时间要完全一样，放大倍数一
样，然后才有可能进行比较。 |
a****e
发帖数: 128 | 3 Thanks for the reply. I did everything exactly the same but still find
intensity variations between the wild type and the mutant sections. My
feeling is that the difference in expression level is small, if there is any
, between the wild type and the mutant. But I am just wondering if there is
there anyway to make a solid conclusion about the expression level
difference. |
a***e
发帖数: 1010 | 4 if anything is subtle and hard to reproduce, I prefer not rely on it. |
n********k
发帖数: 2818 | 5 one way to know whether you shall do it or not, is to check how many people
are doing it in their
studies...BTW, as far as I know, what you are trying to do is very very
tricky, and technically rather demanding,
and several companies were trying to develop ways/reagents to control for
this kind of practice...but a Tech
support told me that not many people does it and for most of the people who
have done it, either they are
super great with microscopy work or it is full of craps...In short, it is
【在 a****e 的大作中提到】
: Thanks for the reply. I did everything exactly the same but still find
: intensity variations between the wild type and the mutant sections. My
: feeling is that the difference in expression level is small, if there is any
: , between the wild type and the mutant. But I am just wondering if there is
: there anyway to make a solid conclusion about the expression level
: difference.
|
a****e
发帖数: 128 | 6 What about in situ-hybridization? Is this a better alternative approach
than IF regarding resolution and reliability. Other alternative ways? |
q******g
发帖数: 3858 | |
