h******y
发帖数: 1374 | 1 求一个好用的探测autophagy的antibody呀 |
m********2
发帖数: 974 | 2 你是要探测一个Autophagy-related gene?还是只要能探测到autophagy就可以了？ |
h******y
发帖数: 1374 | 3 LC3B就可以了 呵呵
【在 m********2 的大作中提到】
: 你是要探测一个Autophagy-related gene?还是只要能探测到autophagy就可以了？
|
m********2
发帖数: 974 | |
j****g
发帖数: 406 | 5 我一直用MBL的，效果不错
【在 h******y 的大作中提到】
: 求一个好用的探测autophagy的antibody呀
|
s*****g
发帖数: 7857 | |
m*******n
发帖数: 387 | |
h******y
发帖数: 1374 | 8 给个cat ＃呀
【在 s*****g 的大作中提到】
: Cell signaling 也可以
|
s*****g
发帖数: 7857 | 9 Cell signaling :
以前用
LC3B Antibody #2775
但它是Polyclonal antibodies
现在有
LC3B (D11) XP™ Rabbit mAb #3868
LC3A (D50G8) XP™ Rabbit mAb #4599
买了其中一个,但还没有用. |
h******y
发帖数: 1374 | 10 你用来做IF还是WB呢？
【在 s*****g 的大作中提到】
: Cell signaling :
: 以前用
: LC3B Antibody #2775
: 但它是Polyclonal antibodies
: 现在有
: LC3B (D11) XP™ Rabbit mAb #3868
: LC3A (D50G8) XP™ Rabbit mAb #4599
: 买了其中一个,但还没有用.
|
k*********a
发帖数: 49 | 11 as far as i know autophagy cannot be determined by one single approach because
each method has its limitation, you have to test multiple markers to convince
people; but yes if you only need see preliminary result whether or not autophagy
is involved, you can try lc3 western but it's very tricky because of basal level
background, and it's highly dependent on what cells you use. a relatively more
convenient way is to use gfp-lc3, write email to some autophagy guy and ask for
construct
【在 h******y 的大作中提到】
: 求一个好用的探测autophagy的antibody呀
|
t*x
发帖数: 1065 | 12 恩，LC-3 II antibody可以用来做western-blot，除了抗体外，lz还可以试试下面方法
来共同支持autophagosome的形成：
1. GFP-LC3转化后可以用来看puncta formation
2. 再有MDC assay有时候也还可以用，但比较tricky
3. 最有说服力的应该是TEM了吧，电镜下可以看到autophagosome或者autolysosome
俺不是做autophagy的，但一个小课题涉及到一些，说错了轻拍啊，呵呵，等牛牛们来
回答好了。 |
h******y
发帖数: 1374 | 13 Thanks a lot. We just infected cells with mCherry-LC3B lentivirus and will
see...
Antibodies from Cell Signaling and Abgent are just not working for us...
because
convince
autophagy
level
more
ask for
【在 k*********a 的大作中提到】
: as far as i know autophagy cannot be determined by one single approach because
: each method has its limitation, you have to test multiple markers to convince
: people; but yes if you only need see preliminary result whether or not autophagy
: is involved, you can try lc3 western but it's very tricky because of basal level
: background, and it's highly dependent on what cells you use. a relatively more
: convenient way is to use gfp-lc3, write email to some autophagy guy and ask for
: construct
|
j****g
发帖数: 406 | 14 还可以在做一个long tern protein degradation assay。 |
