L****S 发帖数: 76 | 1 取决于培养条件,additives 比如prolactin; hydrocortison,insulin,sodium
selenite,beta estradial 的量。
因为
:
MCF7 : ER (+);PR (+);HER2 (-/low)
MDA-MB-231: ER (-);PR (-);HER2 (-)
但是如果只用 FBS,MDA-MB-231 应该比MCF7 快。你还要考虑铺板时的细胞数是否一样。
如果只是想看看是否是MDA-MB-231;MCF7, 可以GOOGLE ATCC 或者
http://icbp.lbl.gov/breastcancer/celllines.php |
|
|
g**********0 发帖数: 180 | 3 I cannot find the cell on ATCC website...... |
|
|
a*****n 发帖数: 2835 | 5 要专门的293T或者专门的一种HeLa
查一下ATCC 和INVITROGEN |
|
R*****w 发帖数: 447 | 6 ankyrin, Thank you very much!
You are right. I checked ATCC. found some HeLa clees are mentioned for suspension culture, such as HeLa-S3, H1HeLa (in media without Ca++)
Most other HeLa cell lines are not mentioned for suspension culture. |
|
k****4 发帖数: 43 | 7 这个跟mean不mean没什么关系。我06年的时候找人要一个质粒,那人说有空帮我
找找。结果4年多了还没见他有空。估计压根就忘了。
鼠造血干细胞。查到文献有些实验
记错了,ATCC没的买,可以再找他
Unfortunately we don't have enough
give some away.In this case, I'll let
,类似CHO,转进去了mouse SCF
给老板,是不是这个学生怕麻烦,自作 |
|
O******e 发帖数: 4845 | 8 很正常的事情。是你在求他,不是他求你。Just be patient or ask for it from
somewhere else.
鼠造血干细胞。查到文献有些实验
记错了,ATCC没的买,可以再找他
Unfortunately we don't have enough
give some away.In this case, I'll let
,类似CHO,转进去了mouse SCF
给老板,是不是这个学生怕麻烦,自作 |
|
f****b 发帖数: 2410 | 9 客观环境也推波住蓝,获诺奖的钱社么就是问一个白人要的荧光物质,那个白人现在在
开出租车生
活,诺奖连提都没有提,现在的人文环境太差了,生物就别提了,
鼠造血
干细胞。查到文献有些实验
记错了,
ATCC没的买,可以再找他
Unfortunately
we don't have enough
then give some away.In this case, I'll let
,类似CHO,
转进去了mouse SCF
给老板,是不
是这个学生怕麻烦,自作 |
|
s****9 发帖数: 932 | 10 It is norm.
I would like to share an experience also. We want a cell line from a group.
They said yes but need to charge us $250 for the cell culture associated
expense plus shipment. We want it badly at that time and paid them $250 in
the end.
鼠造血干细胞。查到文献有些实验
记错了,ATCC没的买,可以再找他
Unfortunately we don't have enough
give some away.In this case, I'll let
,类似CHO,转进去了mouse SCF
给老板,是不是这个学生怕麻烦,自作 |
|
w*****r 发帖数: 2061 | 11 还不错了,有的直接不回复
鼠造血干细胞。查到文献有些实验
记错了,ATCC没的买,可以再找他
Unfortunately we don't have enough
give some away.In this case, I'll let
,类似CHO,转进去了mouse SCF
给老板,是不是这个学生怕麻烦,自作 |
|
b**j 发帖数: 415 | 12 mean is usual, nice is unusual regarding this.
鼠造血干细胞。查到文献有些实验
记错了,ATCC没的买,可以再找他
Unfortunately we don't have enough
give some away.In this case, I'll let
,类似CHO,转进去了mouse SCF
给老板,是不是这个学生怕麻烦,自作 |
|
h**********r 发帖数: 671 | 13 难易程度因细菌而异,有的菌你敲两三个基因就发PNAS,有的你挨个儿敲1000个基因也
许可以发PNAS。
敲除可以用mob型的自杀质粒或者类似于Tn5转座的质粒。DNA导入可以电转也可以二亲
(三亲)结合转导。具体情况具体分析。
还有的可以double crossover把marker切掉,利于下一轮的敲除。
你可以看下这篇paper,Small mobilizable multi-purpose cloning vectors derived
from the Escherichia coli plasmids pK18 and pK19: selection of defined
deletions in the chromosome of Corynebacterium glutumicum.
Gene, 145 (1994) 69-73.
质粒pK18mobsacB在G+,G-细菌用的都很广泛,全序列已知。可以从ATCC买到。
house keeping gene不知道,看是不是条件致死啥的或者营养缺陷型。又不知道你说的
是啥细菌。 |
|
g*****n 发帖数: 241 | 14 我从ATCC买回一个质粒,打算PCR之后subclone到我的载体,但那个基因的5’端从ATG
开始的40个碱基只有三个A/T,GC含量是97%,但整个基因的总GC含量正常。我只好把正
向引物设计在质粒上,最后三个碱基和ATG重合。
我用了phusion polymerase的HF buffer,没有带。换了它的GC buffer,试了不同的退
火温度,结果要么是很多带,要么没有。
请问大家有没有什么建议?
谢谢! |
|
y******y 发帖数: 107 | 15 另外在那里能买呢?ATCC好像挺贵的,一个strain要 $205. |
|
b*****l 发帖数: 9499 | 16 说到生物,还有比 FBS 更让人 ft 的东西么?我的 cell line,就只用 ATCC 的 FBS,
别的都不敢试。另一个组,发现只有某个公司的某个 batch 的 FBS 能使他们的细胞出
现某个关键的 differentiation,于是就把那个 batch 全给买下来了,堆了好几个冰箱
,被我们那里传为笑谈。
,结
种
做样
就要 |
|
C******8 发帖数: 602 | 17 最近一段时间长这个cell总会发现media里脏乎乎的,有一些不知道是deris还是啥,
每次多用1xpbs多洗两遍也不行。。。
倒应该真不是mycoplasma contamination,因为前几天测还是negative的。。。
和以前的subculture也没多大区别啊。。。1200rpm5分钟centrifuge的 |
|
D***a 发帖数: 516 | 18 我也养过,发现这不怎么贴壁,轻轻一吹打就下来。可能就是这样死细胞多才脏吧。 |
|
r**o 发帖数: 212 | 19 给ATCC打电话了,他们也用的是bovine提取的insulin。合成的人胰岛素不适用于我的
这个鼠细胞。
谢谢楼上的回复! |
|
b******n 发帖数: 4225 | 20 今天收到卡片,说是开卖Klebsiella pneumoniae (NDM-1)了
还是很快速的 |
|
s**********e 发帖数: 2888 | 21 我还看过一个组发的一系列的cancer research,报道一个分子在各种癌细胞的
proliferation,就是MTT assay做出来的。我就去重复了,atcc新买的细胞,各种浓度
,时间,serum starvation, collagen coating,mtt, brdu,能试的都试了,什么效果
也看不到。
给那个组发信,没有任何回复。给合作小组写信,合作组说他们也什么都没有看到,而
且那个分子理论上应该被cancer research的cell line很快的降解掉,所以应该是什么
效果也看不到。
6
chemical |
|
|
|
e******0 发帖数: 2149 | 24
多谢! 想再请教下一般你是哪几代做实验?我查了文献差别都很大,我用的是ATCC的
细胞。 |
|
|
n*********1 发帖数: 8 | 26 I got the information from ATCC, but how can we know if the materials have
been tested on rodents or human transmissible pathogens. From the website, i
didn't get it.
Sorry for asking these "stupid" questions. |
|
A******y 发帖数: 2041 | 27 There are a lot of immortalize techniques...for example the most common
human kidney cell line Hek293 is an embryonic kidney cell transformed with
adenovirus 5 DNA. Usually the immortalization is included in the cell line
information sheet.
http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/ta |
|
A******d 发帖数: 571 | 28 I am working with a mouse liver cell line (Hepa1-6) from ATCC. There are
always clumps of cells in the flask here and there. I tried pipeting up and
down- didn't work. I even vortexed the cell suspension- cells are still
clumped.Anyone has experience of breaking up clumped cells? Thanks! |
|
|
h*****t 发帖数: 103 | 30 谢谢,但是有些迷糊了。
因为ATCC上面这样写的:
Description
lymphoblast, established from long-term bone marrow cultures of C3H/HeJ mice
infected with the Friend murine leukemia virus. Morphology: mostly round
cells growing singly or in small clusters in suspension, some cells are
loosely adherent.
但是,另外一个人的paper却写的该细胞为myeloid:
Shi B, Prisco M, Calin G, Liu CG, Russo G, Giordano A, Baserga R. Expression
profiles of micro RNA in proliferating and differentiating 32D murine
myeloid cells. J Cell Physiology 207: 706-10,... 阅读全帖 |
|
|
|
l****3 发帖数: 399 | 33 倒个有抗生素的板子,一划不就知道了吗
或者梯度的液体培养,看看多高浓度有抑制 |
|
H*****e 发帖数: 120 | 34 1. H1299 needs 2-3 weeks for seeing colonies, whereas H460 colonies can be
seen in 5-7 day. If it still does not work, change the mediumn every 4 days
or order new cells from ATCC. We routinely do it and no problem found.
2.It has nothing to do with stem cells. Most stem cell medium are serum
free. But Soft agar uses 10-20% serum (I even saw people use 30% serum for
weak oncogene transformed 3T3.).
Good luck! |
|
l***y 发帖数: 638 | 35 search IMAGE clones, then order from ATCC, Thermo/Fisher, Invitrogen |
|
a****l 发帖数: 125 | 36 Origene
ATCC
Invitrogen
Open Biosystems |
|
m****M 发帖数: 360 | 37 ATCC has the set of Adeasy system for only around $250 |
|
i*****g 发帖数: 11893 | 38 简单说,用药是杀不光的,无论说明书多好多好
对于要长期使用的细胞系,唯一的办法是不感染mycoplasma
还有,据说某些ATCC的细胞系,时间一长就有myco,虽然开始时候检测不到 |
|
m****g 发帖数: 42 | 39 NK92 是NK tumor cell line, you can buy it from ATCC. 看来你们不想跟人合作,
这也可以理解,特别是有望成为药物的东西。要是有钱,没什么作不成的。生物方面的
postdoc还是相对容易找到的。 |
|
l**********1 发帖数: 5204 | 40 //www.ncbi.nlm.nih.gov/pubmed/21050804
浮黴菌門
Isosphaera屬
//hamap.expasy.org/proteomes/ISOPI.html
Species: Isosphaera pallida (strain ATCC 43644 / DSM 9630 / IS1B)
Species code: ISOPI
Taxonomy: Bacteria; Planctomycetes; Planctomycetia; Planctomycetales; Planctomycetaceae; Isosphaera
(TaxID: 575540) [NEWT/ NCBI]
Description: Isosphaera pallida strain IS1B was isolated from a hot spring in Kah-nee-tah, Oregon, USA.
It is the only budding bacterium known to glide and the only heterotrophic bacteriu... 阅读全帖 |
|
l**********1 发帖数: 5204 | 41 别说真核生物的感光系统的细胞能利用光 通信
就是浮黴菌門Isosphaera屬
的细菌Isosphaera pallida 说不定也能利用光 通信呢
Isosphaera pallida strain IS1B was isolated from a hot spring in Kah-nee-tah
, Oregon, USA.
It is the only budding bacterium known to glide and the only heterotrophic
bacterium known to be phototactic
Species: Isosphaera pallida (strain ATCC 43644 / DSM 9630 / IS1B)
Species code: ISOPI
Taxonomy: Bacteria; Planctomycetes; Planctomycetia; Planctomycetales;
Planctomycetaceae; Isosphaera
(TaxID: 575540) [NEWT/ NCB... 阅读全帖 |
|
|
g***y 发帖数: 201 | 43 我拍了很多图片发给他们了,貌似他们在调查。在这里提醒大家ATCC的细胞也不是那么
可靠的。
“The Specialist has authorized a growing flask。。。。 This will allow us
to start another culture from a frozen vial and observe it for visual
contamination. ” |
|
b***2 发帖数: 348 | 44 请教一下做ips的大牛们。
我主要是研究大分子投递的。因为ips很火,并且做的方法也算是有限,病毒效果好,
但是有风险,其他的方法据说效果差,我现在想设计一种方法,不用病毒,期望达到接
近病毒的效果。
现在的问题是我没有杀老鼠的protocol,这样自己搞MEF有难度,ATCC的又贵,现在我
的方法也不一定能成功,老板不见兔子不愿意撒鹰出去。我想问一下,初始的细胞材料
,包括做reprogram的和feeder有没有什么容易一些的途径搞到。 |
|
v*******n 发帖数: 8995 | 45 我们隔壁实验室有一个在表达lif 的CHO
用他的condition media就行了。你可以看看atcc有没有卖 |
|
|
l**x 发帖数: 52 | 47 现在用的是C2C12细胞,经常一不小心就长满整个平板,ATCC说这个细胞株不能长满平
板,另外,这个细胞表达myosin regulatory light chain的磷酸化特别低,很难用
WESTERN测出来,用的是Cell Signaling的regulatory light chain S19的磷酸化抗体。
大家有什么好的细胞株推荐吗?特别是来自于skeletal muscle的。另外,对抗体有什
么推荐的最好了,谢谢 |
|
n********k 发帖数: 2818 | 48 C2C12 is both a very easy and very hard line to work with and I am not aware
any other better established line other than this one or one could go with
primary lines...
Anyhow, If one follows closely certain routines, this line is very easy--
-passage before 80-90% confluence with no local overgrowth...Overgrowth is a
huge problem with this line, after 3-5 times, the entire culture will be
changed...Also, the confluence etc matters a lot for differentiation...
In addition, there are at lea... 阅读全帖 |
|
n*********o 发帖数: 210 | 49 rt,实验需要小鼠的肠道上皮细胞系, 最好是正常小鼠的colon epithelial cell
line. 可能由于正常小鼠的细胞难于培养或存活吧,我在ATCC没能找到。有用过的或见
过的请帮忙介绍下。谢谢! |
|
|