z*******6
发帖数: 679 | 1 号称这种小的inhibitor很快就会被clear... 大家有没有用过的？目前就找到一篇J
Neuro i.p. injection了，其他好像没见过system administration... 希望板上同学
给出出主意...
我想用in vivo的model看NO对endothelial adhesion molecules的调控... 因为in
vitro都被人做了，我的优势也就是有个特殊的in vivo model... |
z*****n
发帖数: 444 | 2 Lnna,放水里，好像不是很好溶解，配stock，调碱性。 |
n***w
发帖数: 2405 | 3 you can dissolve in PBS and I think the solubility is up to 10mg/ml in PBS,
if I remember correctly. You can also check the package insert for L-NAME or
the MSDS to find such information.
I make mine at 5mg/ml in PBS and the dose is 30mg/kg body weight.
The half-time of L-NAME varies in literature but the ballpark is around
250min to 300min.
I do i.p. injection and do my experiments after 2 hours. I see an effective
lowering of Nitrate+Nitrite level. Some papers even do it after 1 hour.
If you want to long-term experiment, I guess you have to mix with drinking
water. You can find whole bunch of papers doing this way. |
z*******6
发帖数: 679 | 4 我给老鼠打了3.5小时以后做qPCR检测我的molecule表达... 对了，我用D-NAME做的
control，应该可以吧？
结果发现i.p.和i.v.打结果不一样。。。
我是按mole算的，不是按g...
iv打了90ul 100mM在PBS里
ip打了360ul 100mM也在PBS里
结果发现iv打的positive control出来了，结果ip没出来... 我不打算纠结到底为啥ip
没出来，就打算重复一两次iv，然后positive control结果一致就打算用它了...
,
or
effective
【在 n***w 的大作中提到】
: you can dissolve in PBS and I think the solubility is up to 10mg/ml in PBS,
: if I remember correctly. You can also check the package insert for L-NAME or
: the MSDS to find such information.
: I make mine at 5mg/ml in PBS and the dose is 30mg/kg body weight.
: The half-time of L-NAME varies in literature but the ballpark is around
: 250min to 300min.
: I do i.p. injection and do my experiments after 2 hours. I see an effective
: lowering of Nitrate+Nitrite level. Some papers even do it after 1 hour.
: If you want to long-term experiment, I guess you have to mix with drinking
: water. You can find whole bunch of papers doing this way.
|
n***w
发帖数: 2405 | 5 你指的positive control是啥？
ip
【在 z*******6 的大作中提到】
: 我给老鼠打了3.5小时以后做qPCR检测我的molecule表达... 对了，我用D-NAME做的
: control，应该可以吧？
: 结果发现i.p.和i.v.打结果不一样。。。
: 我是按mole算的，不是按g...
: iv打了90ul 100mM在PBS里
: ip打了360ul 100mM也在PBS里
: 结果发现iv打的positive control出来了，结果ip没出来... 我不打算纠结到底为啥ip
: 没出来，就打算重复一两次iv，然后positive control结果一致就打算用它了...
:
: ,
|
z*******6
发帖数: 679 | 6 E-selectin上调了... 按照文献抑制NO可以激活NF-kB pathway，从而上调E-selectin
，VCAM-1，ICAM-1之流...
【在 n***w 的大作中提到】
: 你指的positive control是啥？
:
: ip
|
n***w
发帖数: 2405 | 7 额，这一切都在3.5小时发生？
你还是测一下nitrite/nitrate比较靠谱。也许你两种注射方式都抑制了nos，只是程度
不一样导致了你的readout差异。
另外，你这个ip的剂量有依据吗？360ul感觉好多阿。。。
selectin
【在 z*******6 的大作中提到】
: E-selectin上调了... 按照文献抑制NO可以激活NF-kB pathway，从而上调E-selectin
: ，VCAM-1，ICAM-1之流...
|
