求助:有什么方法能boost cytoplasmic calcium concentration in MEFs?# Biology - 生物学
r*n
1 楼
想退掉或换新的，买了一个多月了，
包装盒都给扔了，
请问怎么操作。
包装盒都给扔了，
请问怎么操作。
g*t
2 楼
我假装自己是java老师傅，一点馅也没漏。诀窍是，每当提起python 时，都摇头叹气
加鄙视就可以了。哈哈。
Java 社务人多心齐，我看好。唯一可虑者就是内贼goog android, mess up java
ecosystem.
加鄙视就可以了。哈哈。
Java 社务人多心齐，我看好。唯一可虑者就是内贼goog android, mess up java
ecosystem.
E*1
3 楼
请问一下各位都知道哪些方法可以提高胞内Ca2+浓度?我用的是MEFs.
我现在只知道一种是用50mM的KCl加到culture medium里面，induce depolarization,
然后使voltage-dependent calcium channel打开，使胞内Ca2+升高。但是有一点不确
定的是MEFs是否有voltage-dependent calcium channel。
求各位指点，在此先谢过了！~
我现在只知道一种是用50mM的KCl加到culture medium里面，induce depolarization,
然后使voltage-dependent calcium channel打开，使胞内Ca2+升高。但是有一点不确
定的是MEFs是否有voltage-dependent calcium channel。
求各位指点，在此先谢过了！~
N*S
4 楼
找厂家，一般都是lifetime的warranty的
f*2
5 楼
golang老中多，java社区咖喱味重
n*w
6 楼
add ATP or adenosine.
It will activate adenosine receptor, converting PIP2 to IP3 which binds to
IP3 receptor on ER to liberate calcium.
It will activate adenosine receptor, converting PIP2 to IP3 which binds to
IP3 receptor on ER to liberate calcium.
s*i
7 楼
从新插一下就好了。
c*n
8 楼
话说vertx这玩意真是曲高和寡吗
账面上看着很不错 但是实在没看到太多用的人 总感觉是个大坑
【在 f******2 的大作中提到】
: golang老中多，java社区咖喱味重
账面上看着很不错 但是实在没看到太多用的人 总感觉是个大坑
【在 f******2 的大作中提到】
: golang老中多，java社区咖喱味重
E*1
9 楼
Thanks a lot!
I am wondering whether it is a general method, which means this method also
works on MEFs.
And I need more details about the method you mentioned, so could you give me
a link of the protocol or a paper where the method has been used.
【在 n***w 的大作中提到】
: add ATP or adenosine.
: It will activate adenosine receptor, converting PIP2 to IP3 which binds to
: IP3 receptor on ER to liberate calcium.
r*n
10 楼
重新插了三遍了
memtest86+ 依然报错
【在 s********i 的大作中提到】
: 从新插一下就好了。
memtest86+ 依然报错
【在 s********i 的大作中提到】
: 从新插一下就好了。
g*t
11 楼
烙印可以合作。至少我觉得轻松。那些小心思我觉得很cute，可爱，幼稚。
而且老印整体素质比老中高。实话实说，如果没有烙印整的这个IT圈子。中国马工
的工作机会会少很多。
再实话实说，我整不过白人里面那种本科烂校，年纪轻轻
升到大公司SVP的所谓alpha male。能整过现在也不是正确的时机。
【在 f******2 的大作中提到】
: golang老中多，java社区咖喱味重
而且老印整体素质比老中高。实话实说，如果没有烙印整的这个IT圈子。中国马工
的工作机会会少很多。
再实话实说，我整不过白人里面那种本科烂校，年纪轻轻
升到大公司SVP的所谓alpha male。能整过现在也不是正确的时机。
【在 f******2 的大作中提到】
: golang老中多，java社区咖喱味重
n*w
12 楼
It works on MEFs. Please refer to the forwarded article:
http://www.ncbi.nlm.nih.gov/pubmed/21084727
also
me
【在 E**********1 的大作中提到】
:
: Thanks a lot!
: I am wondering whether it is a general method, which means this method also
: works on MEFs.
: And I need more details about the method you mentioned, so could you give me
: a link of the protocol or a paper where the method has been used.
http://www.ncbi.nlm.nih.gov/pubmed/21084727
also
me
【在 E**********1 的大作中提到】
:
: Thanks a lot!
: I am wondering whether it is a general method, which means this method also
: works on MEFs.
: And I need more details about the method you mentioned, so could you give me
: a link of the protocol or a paper where the method has been used.
r*n
13 楼
刚买了两条，
如果新的没有问题，就说明旧的确实有问题，
然后去找厂家试试，看是能退，还是换
【在 N*****S 的大作中提到】
: 找厂家，一般都是lifetime的warranty的
如果新的没有问题，就说明旧的确实有问题，
然后去找厂家试试，看是能退，还是换
【在 N*****S 的大作中提到】
: 找厂家，一般都是lifetime的warranty的
E*1
14 楼
I've checked that paper. Based on my understanding, the paper said ATP
stimulation can temporarily inhibit respiration and reduce mitochondrial
membrane potential, so inhibit mitochondrial Ca2+ uptake. After temporary
inhibition of Ca2+ uptake, ATP is consumed and mitochondrial respiration
resumed and cause mitochondrial Ca2+ boost. And the boost after ATP
stimulation can reflect the ability of cellular respiration.
But this still doesn't solve my problem, which needs to increase cytoplasmic
Ca2+ level.
【在 n***w 的大作中提到】
: It works on MEFs. Please refer to the forwarded article:
: http://www.ncbi.nlm.nih.gov/pubmed/21084727
:
: also
: me
d*t
15 楼
超过30天不给退了吧，只好去找厂家RMA
【在 r*******n 的大作中提到】
: 刚买了两条，
: 如果新的没有问题，就说明旧的确实有问题，
: 然后去找厂家试试，看是能退，还是换
【在 r*******n 的大作中提到】
: 刚买了两条，
: 如果新的没有问题，就说明旧的确实有问题，
: 然后去找厂家试试，看是能退，还是换
n*w
16 楼
No. You misunderstood the paper. ATP is used to activate the adenosine
receptor.
The largest calcium store inside the cell is endoplasmic reticulum, which
can release a large amount of calcium upon IP3-R is activated.
Mitochondria is critical in maintaining calcium homeostasis and will take up
calcium if there is calcium overload.
You can transfect your MEFs with cytoplasmic aequorin (you can search on
this to get a better idea), which is widely used as a intracellular calcium
indicator.
You can check on this paper also: PMID 21720385, look at Figure 3 C and the methods section. But they did
not perform experiments on MEFs.
Good luck.
cytoplasmic
【在 E**********1 的大作中提到】
:
: I've checked that paper. Based on my understanding, the paper said ATP
: stimulation can temporarily inhibit respiration and reduce mitochondrial
: membrane potential, so inhibit mitochondrial Ca2+ uptake. After temporary
: inhibition of Ca2+ uptake, ATP is consumed and mitochondrial respiration
: resumed and cause mitochondrial Ca2+ boost. And the boost after ATP
: stimulation can reflect the ability of cellular respiration.
: But this still doesn't solve my problem, which needs to increase cytoplasmic
: Ca2+ level.
receptor.
The largest calcium store inside the cell is endoplasmic reticulum, which
can release a large amount of calcium upon IP3-R is activated.
Mitochondria is critical in maintaining calcium homeostasis and will take up
calcium if there is calcium overload.
You can transfect your MEFs with cytoplasmic aequorin (you can search on
this to get a better idea), which is widely used as a intracellular calcium
indicator.
You can check on this paper also: PMID 21720385, look at Figure 3 C and the methods section. But they did
not perform experiments on MEFs.
Good luck.
cytoplasmic
【在 E**********1 的大作中提到】
:
: I've checked that paper. Based on my understanding, the paper said ATP
: stimulation can temporarily inhibit respiration and reduce mitochondrial
: membrane potential, so inhibit mitochondrial Ca2+ uptake. After temporary
: inhibition of Ca2+ uptake, ATP is consumed and mitochondrial respiration
: resumed and cause mitochondrial Ca2+ boost. And the boost after ATP
: stimulation can reflect the ability of cellular respiration.
: But this still doesn't solve my problem, which needs to increase cytoplasmic
: Ca2+ level.
s*y
17 楼
大赞
up
calcium
the methods section. But they did
【在 n***w 的大作中提到】
: No. You misunderstood the paper. ATP is used to activate the adenosine
: receptor.
: The largest calcium store inside the cell is endoplasmic reticulum, which
: can release a large amount of calcium upon IP3-R is activated.
: Mitochondria is critical in maintaining calcium homeostasis and will take up
: calcium if there is calcium overload.
: You can transfect your MEFs with cytoplasmic aequorin (you can search on
: this to get a better idea), which is widely used as a intracellular calcium
: indicator.
: You can check on this paper also: PMID 21720385, look at Figure 3 C and the methods section. But they did
up
calcium
the methods section. But they did
【在 n***w 的大作中提到】
: No. You misunderstood the paper. ATP is used to activate the adenosine
: receptor.
: The largest calcium store inside the cell is endoplasmic reticulum, which
: can release a large amount of calcium upon IP3-R is activated.
: Mitochondria is critical in maintaining calcium homeostasis and will take up
: calcium if there is calcium overload.
: You can transfect your MEFs with cytoplasmic aequorin (you can search on
: this to get a better idea), which is widely used as a intracellular calcium
: indicator.
: You can check on this paper also: PMID 21720385, look at Figure 3 C and the methods section. But they did
E*1
18 楼
Really thank you!
Sorry about my misunderstanding. I really don't know much about calcium.
You mentioned use aequorin as a calcium indicator. My question is how about
Fura2-AM. What are the pros and cons of these two? After the treatment, I
will do immunoflurescence staining, I am thinking if I use Fura2-AM, I can
get calcium level and localization of my target protein at the same time.
Really thanks for you time!
up
calcium
the methods section. But they did
【在 n***w 的大作中提到】
: No. You misunderstood the paper. ATP is used to activate the adenosine
: receptor.
: The largest calcium store inside the cell is endoplasmic reticulum, which
: can release a large amount of calcium upon IP3-R is activated.
: Mitochondria is critical in maintaining calcium homeostasis and will take up
: calcium if there is calcium overload.
: You can transfect your MEFs with cytoplasmic aequorin (you can search on
: this to get a better idea), which is widely used as a intracellular calcium
: indicator.
: You can check on this paper also: PMID 21720385, look at Figure 3 C and the methods section. But they did
Sorry about my misunderstanding. I really don't know much about calcium.
You mentioned use aequorin as a calcium indicator. My question is how about
Fura2-AM. What are the pros and cons of these two? After the treatment, I
will do immunoflurescence staining, I am thinking if I use Fura2-AM, I can
get calcium level and localization of my target protein at the same time.
Really thanks for you time!
up
calcium
the methods section. But they did
【在 n***w 的大作中提到】
: No. You misunderstood the paper. ATP is used to activate the adenosine
: receptor.
: The largest calcium store inside the cell is endoplasmic reticulum, which
: can release a large amount of calcium upon IP3-R is activated.
: Mitochondria is critical in maintaining calcium homeostasis and will take up
: calcium if there is calcium overload.
: You can transfect your MEFs with cytoplasmic aequorin (you can search on
: this to get a better idea), which is widely used as a intracellular calcium
: indicator.
: You can check on this paper also: PMID 21720385, look at Figure 3 C and the methods section. But they did
n*w
19 楼
I do not know much about Fura2-AM. I read papers that use this dye
in neurons but not in MEFs.
Based on your objective, I do think probably using a dye may be better than
aequorin. But the experimental conditions need to be carefully taken care of
since Fura2-AM imaging may be affected by many variables.
Good luck.
about
【在 E**********1 的大作中提到】
: Really thank you!
: Sorry about my misunderstanding. I really don't know much about calcium.
: You mentioned use aequorin as a calcium indicator. My question is how about
: Fura2-AM. What are the pros and cons of these two? After the treatment, I
: will do immunoflurescence staining, I am thinking if I use Fura2-AM, I can
: get calcium level and localization of my target protein at the same time.
: Really thanks for you time!
:
: up
: calcium
in neurons but not in MEFs.
Based on your objective, I do think probably using a dye may be better than
aequorin. But the experimental conditions need to be carefully taken care of
since Fura2-AM imaging may be affected by many variables.
Good luck.
about
【在 E**********1 的大作中提到】
: Really thank you!
: Sorry about my misunderstanding. I really don't know much about calcium.
: You mentioned use aequorin as a calcium indicator. My question is how about
: Fura2-AM. What are the pros and cons of these two? After the treatment, I
: will do immunoflurescence staining, I am thinking if I use Fura2-AM, I can
: get calcium level and localization of my target protein at the same time.
: Really thanks for you time!
:
: up
: calcium
E*1
20 楼
than
of
Thanks a ton, bro!
【在 n***w 的大作中提到】
: I do not know much about Fura2-AM. I read papers that use this dye
: in neurons but not in MEFs.
: Based on your objective, I do think probably using a dye may be better than
: aequorin. But the experimental conditions need to be carefully taken care of
: since Fura2-AM imaging may be affected by many variables.
: Good luck.
:
: about
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