a****l
发帖数: 125 | 1 western blotting 有时定量不精确，不太大的变化有是看不出来。有什么更敏感的方
法呢？
谢谢 |
r***e
发帖数: 2539 | 2 ELISA?
【在 a****l 的大作中提到】
: western blotting 有时定量不精确，不太大的变化有是看不出来。有什么更敏感的方
: 法呢？
: 谢谢
|
g*****y
发帖数: 34 | |
k******0
发帖数: 1073 | 4 radiolabelling ?
Quantitative MS |
A******y
发帖数: 2041 | 5 If you cannot see the difference in Western, is it really relevant
biologically? Yes, ELISA, Alpha-scan, and quantitative MS...how about
iodinated protein A quantization... |
s******y
发帖数: 28562 | 6 话说回来ELISA也是用抗体来做的，为什么比western 敏感？
【在 r***e 的大作中提到】
: ELISA?
|
w******e
发帖数: 1187 | 7 因为sensitivity跟signal amplification有关。
btw WB的LOD通常是多少？pM还是nM？
【在 s******y 的大作中提到】
: 话说回来ELISA也是用抗体来做的，为什么比western 敏感？
|
s******y
发帖数: 28562 | 8 最低可以达到femto-gram
【在 w******e 的大作中提到】
: 因为sensitivity跟signal amplification有关。
: btw WB的LOD通常是多少？pM还是nM？
|
w******e
发帖数: 1187 | 9 差这么多啊。。。以前做WB的时候都没意识到这个问题。。。
nb的ELISA<1pM LOD很正常的
【在 s******y 的大作中提到】
: 最低可以达到femto-gram
|
s******y
发帖数: 28562 | 10 啊，我好像算错数了。应该是除法我算成乘法了。抱歉抱歉。
天色太晚，我头都晕了。还是赶快睡觉去了。呵呵 |
|
|
m******g
发帖数: 69 | 11 ELISA假阳性太高了。western blot唯一的好处就是那个SDS-PAGE步骤。把蛋白按按照
质量分离了，从而部分消除了抗体非特异性可能造成的假阳性。可是ELISA只能是测定
抗体抗原结合程度，但是抗体是否特异性结合抗原，无法得知。但是ELISA如果做好了
，比WB好敏感多了，可以做error bar，也可以做到pg水平。 |
A******y
发帖数: 2041 | 12 I thought the ability for western blot to tell the difference has to do with
the linearity of the signal and maximum signal to noise ratio. If you use
film, you are limited by the film's linearity range which I "heard" is
between 2-10, so you need at least two fold differences to see a different.
Using a digital imager, some claim to have x2000 to x1000 fold linear range
especially if you are using fluorescent probes.
ELSA has high false rates that's why alpha-scan is developed. |
A******y
发帖数: 2041 | 13 I thought the ability for western blot to tell the difference has to do with
the linearity of the signal and maximum signal to noise ratio. If you use
film, you are limited by the film's linearity range which I "heard" is
between 2-10, so you need at least two fold differences to see a different.
Using a digital imager, some claim to have x2000 to x1000 fold linear range
especially if you are using fluorescent probes.
ELSA has high false rates that's why alpha-scan is developed. |
a****l
发帖数: 125 | 14 没太懂digital imager part, x2000 和 x1000 是什么意思？是比film 敏感 1000－
2000 倍？
with
use
.
range
【在 A******y 的大作中提到】
: I thought the ability for western blot to tell the difference has to do with
: the linearity of the signal and maximum signal to noise ratio. If you use
: film, you are limited by the film's linearity range which I "heard" is
: between 2-10, so you need at least two fold differences to see a different.
: Using a digital imager, some claim to have x2000 to x1000 fold linear range
: especially if you are using fluorescent probes.
: ELSA has high false rates that's why alpha-scan is developed.
|
g*********5
发帖数: 2533 | 15 定量是licor的长处。因为是二抗直接带的荧光，不需要显影，
线形很好。
with
use
.
range
【在 A******y 的大作中提到】
: I thought the ability for western blot to tell the difference has to do with
: the linearity of the signal and maximum signal to noise ratio. If you use
: film, you are limited by the film's linearity range which I "heard" is
: between 2-10, so you need at least two fold differences to see a different.
: Using a digital imager, some claim to have x2000 to x1000 fold linear range
: especially if you are using fluorescent probes.
: ELSA has high false rates that's why alpha-scan is developed.
|
