j**W
发帖数: 89 | | y******8
发帖数: 1764 | 2 There are lots of P-Ser antibodies, but none of them are good.
P-32 works better if you are not afraid of radioactive. | e****s
发帖数: 1125 | 3 Try Cell signaling. Never use anti-p-Ser, but I used their anti-PTyr and
anti-PThr before, and both are good. | z********i
发帖数: 610 | 4 可以试试zymed的pSer（现在是invitrogene），purified的多抗，运气好的话能找到一
个好用的。我们买过好几只，其中有很好用的。
sigma的pSer是单抗，信号很弱可以用。
pY很好用，那是因为都是同一个单克隆细胞株，不同的公司名称可能不一样。
【在 e****s 的大作中提到】
: Try Cell signaling. Never use anti-p-Ser, but I used their anti-PTyr and
: anti-PThr before, and both are good.
| y******8
发帖数: 1764 | 5 pSer Ab does not work well because there are lots of extracellular pSer
signal. If the immune system develops a pSer only antibody, it will be a
huge problem for animal itself.
Extracellular pThr is much less abundant than pSer, so pThr is OK to use.
There is very little extracellular pTyr in physiological condition, and pTyr
Ab works the best.
【在 z********i 的大作中提到】
: 可以试试zymed的pSer（现在是invitrogene），purified的多抗，运气好的话能找到一
: 个好用的。我们买过好几只，其中有很好用的。
: sigma的pSer是单抗，信号很弱可以用。
: pY很好用，那是因为都是同一个单克隆细胞株，不同的公司名称可能不一样。
| j**W
发帖数: 89 | 6 I sort of understand that it is hard to recognize pan pSer because there are
so many of them. However, I am not sure what does it to do with
extracellular localization if the sample is total cell extract.We made some
S->A mutations at a site of interest and observed difference. I just hope to
test whether and how much wild type is phosphorylated at that Ser site with
certain treatments, without making a specific pSer motif antibody ourselves
.
BTW, is there a good program to predict what kinase it may be for a specific
site? I used a couple of web programs and cannot tell what kinase site it
may be.
pTyr
【在 y******8 的大作中提到】
: pSer Ab does not work well because there are lots of extracellular pSer
: signal. If the immune system develops a pSer only antibody, it will be a
: huge problem for animal itself.
: Extracellular pThr is much less abundant than pSer, so pThr is OK to use.
: There is very little extracellular pTyr in physiological condition, and pTyr
: Ab works the best.
| j**W
发帖数: 89 | 7 Does zymed have several pSer antibodies? Will you please let me know which
one is good. Thanks!
【在 z********i 的大作中提到】
: 可以试试zymed的pSer（现在是invitrogene），purified的多抗，运气好的话能找到一
: 个好用的。我们买过好几只，其中有很好用的。
: sigma的pSer是单抗，信号很弱可以用。
: pY很好用，那是因为都是同一个单克隆细胞株，不同的公司名称可能不一样。
| y******8
发帖数: 1764 | 8 Anything extracellular is a potential antigen. Pure pSer antibody in the
animal would develop autoimmune diseases.
So, the pSer antibodies on market are not agaist pure pSer, and are always
context dependent.
If you have a clear target protein, just develop a site specific antibody.
Other wise, p32 is not bad to try since majority of phospho signal comes
from pSer.
are
some
to
with
ourselves
specific
【在 j**W 的大作中提到】
: I sort of understand that it is hard to recognize pan pSer because there are
: so many of them. However, I am not sure what does it to do with
: extracellular localization if the sample is total cell extract.We made some
: S->A mutations at a site of interest and observed difference. I just hope to
: test whether and how much wild type is phosphorylated at that Ser site with
: certain treatments, without making a specific pSer motif antibody ourselves
: .
: BTW, is there a good program to predict what kinase it may be for a specific
: site? I used a couple of web programs and cannot tell what kinase site it
: may be.
| y******8
发帖数: 1764 | 9 For the phospho-site prediction, unless your kinase is defined, it is very
hard. Some kinase has very conserved target sequence, but others display no
pattern.
are
some
to
with
ourselves
specific
【在 j**W 的大作中提到】
: I sort of understand that it is hard to recognize pan pSer because there are
: so many of them. However, I am not sure what does it to do with
: extracellular localization if the sample is total cell extract.We made some
: S->A mutations at a site of interest and observed difference. I just hope to
: test whether and how much wild type is phosphorylated at that Ser site with
: certain treatments, without making a specific pSer motif antibody ourselves
: .
: BTW, is there a good program to predict what kinase it may be for a specific
: site? I used a couple of web programs and cannot tell what kinase site it
: may be.
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