It's an ER-resident protein. According to the literature, upon kinase stimulation, it goes inside the nucleus. We did see nucleus localization for a slightly-truncated version. We have a hypothesis how this happens based on other data we have, and is confirming this using cell-based assay. Would not reveal identity of this protein for now, but it is up-regulated in two types of endocrinological cancers.
【在 m******5 的大作中提到】 : could you share more information regarding your protein?
to verify if you Ab is good for ChIP: Follow your chip protocol to where you precipitate with beads(agarose or magnetic), DO NOT decrosslink, directly do a western blot.
【在 C*********m 的大作中提到】 : Commercially available. So far so good for imaging-based localization : experiment compared to epitope-antibody.
This may not be enough. I did the same thing before for ChIP-qPCR before. Non-specific banding is a major issue; that is the reason why so many emphasize the "good" antibody here.
【在 a******r 的大作中提到】 : to verify if you Ab is good for ChIP: : Follow your chip protocol to where you precipitate with beads(agarose or : magnetic), DO NOT decrosslink, directly do a western blot.
h*o
33 楼
高妹找了好几个帅哥教授了, 这真的是小草。
m*5
34 楼
in such case, I guess it would be sufficient to just include a 'no epitop' group for Chip-Seq.Do peak calling against both input and non epitop.
a
【在 t**********n 的大作中提到】 : This may not be enough. : I did the same thing before for ChIP-qPCR before. Non-specific banding is a : major issue; that is the reason why so many emphasize the "good" antibody : here.