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h******3
发帖数: 3 | 2 Req ID: 383BR
PhD in Chemical Engineering, Biochemistry or a related field with 0-2 years
of experience...
Protein purification, protein chromatography (i.e.GE AKTAs...), filtration
(i.e. TFF...) and related protein analytical techniques.
http://careers.regeneron.com/rensselaer/other/process-scientist |
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R******d
发帖数: 1305 | 3 嗯，我今天实在protein不够量，吃了一个含30g protein的protein bar |
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n*****e
发帖数: 40 | 4 【 以下文字转载自 Biology 讨论区 】
发信人: napaene (北极熊妈妈), 信区: Biology
标 题: 发个工作广告Protein Scientist， Ann Arbor (转载)
发信站: BBS 未名空间站 (Fri Apr 1 10:31:36 2016, 美东)
发信人: napaene (北极熊妈妈), 信区: Michigan
标 题: 发个工作广告Protein Scientist， Ann Arbor
关键字: 生物工作
发信站: BBS 未名空间站 (Fri Apr 1 10:28:57 2016, 美东)
公司在招工，需要protein and antibody 经验。Job post:
http://www.enzolifesciences.com/about-us/careers/product-manufa
需要更多info可以站内联系。 |
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U**********b
发帖数: 13 | 5 【 以下文字转载自 Chemistry 讨论区 】
发信人: UofLlaserlab (UofLlaserlab), 信区: Chemistry
标 题: postdoctoral researcher in solution NMR for protein conformational dynamics
发信站: BBS 未名空间站 (Sat Jan 21 19:01:07 2017, 美东)
The Li lab in the Department of Chemistry at University of Louisville is
seeking a highly motivated postdoctoral researcher to join our lab
immediately.
Our research interests focus on applying solution NMR techniques to
understand the functional roles of protein conformational dynamics in the
deu... 阅读全帖 |
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s***y
发帖数: 24 | 6 【 以下文字转载自 Immigration 讨论区 】
发信人: sevny (forever), 信区: Immigration
标 题: guest editor 机会，protein interaction
发信站: BBS 未名空间站 (Thu Aug 28 21:32:49 2014, 美东)
guest editor机会.
Journal: Biochemistry insights journal supplement (Libertas Academica)
Scope: Protein binding and protein interactions
Tasks: 1. Solicit five papers
2. Compose an editorial
如果有兴趣，请站内联系。 |
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K*F
发帖数: 120 | 7 为申请EB1-A，求审稿机会，Biochemistry, protein chemistry, protein
engineering方面，非常感谢！ |
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d****t
发帖数: 1284 | 8 i like it, i think it tastes better than ON Whey.
it's not pure whey protein, it has a sustained-release formula. also, it has
more calories per serving than whey protein. if you are calories sensitive,
you shouldn't use it.
i do use whey protein after work out for the quick absorption instead of
using BSN though. |
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j********6
发帖数: 207 | 9 请问GNC的whey protein和muscle milk有什么区别？哪个好？
我一直用GNC的whey protein， 昨天听有人说muscle milk比whey protein好，请问这
两个具体有什么区别？谢谢 |
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l****q
发帖数: 767 | 10 一位师姐说是因为nuts是protein 的一种很好的来源，所以划为protein那一类。
划为protein，不是老师划的，是美国那什么金字塔食物分类 |
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s*******t
发帖数: 7746 | 11 哈哈，这些文章看看就罢了。他们定义>20%的热量来自protein就算是high protein了
。生物研究so far就是盲人摸象，怎么摸怎么像，连研究作者也说了：
“People will say, ‘Here we go again: First you attack the fats, then you
attack the carbohydrates, now it’s the protein,’ ” Longo said.
结论是什么都不能吃了：）或者说什么都可以吃，不过量就好了。 |
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f*******r
发帖数: 71 | 12 我一直想减肥，看了老狼的《七年之痒》，知道了nancy clark's sports nutrition
guidebook。 那本书上说很少有人需要人工的蛋白，实际上过多的蛋白没有好处。
看到网上大牛们用这个，我又迷惘了。
Page 225：
Few athletes need to spend money on protein supplements. Even
vegetarians can get enough protein through foods. But sometimes, with
high school and collegiate athletes, as well as triathletes and others doing
double workouts, the need for calories can outweigh the ability to easily
consume them because of lack of appetite or time. And at other times, a
supplement is just mor... 阅读全帖 |
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s******y
发帖数: 28562 | 14 In my own experience, GST is not a good tag because such reason:
1. the GST is on the C term, so , even the protein is not completely
expressed, the GST is still there.
2. GST has a tendency to form dimer (just like what others said)
In your case, it is very likely your protein wasn't expressed correctly.
You can try to induce it by a lower dose of IPTG. In our lab's experience,
sometime it helps to express big proteins. It is also a possibility that you
may introduce a stop codon inside your pr |
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t********m
发帖数: 2 | 15 yes, 4oC o/n or 37oC 5,30,60 and 120min. If you do in column digestion, you
need more enzyme and your protein sample will be diluted, but you do not have
to remove the GST. To my experience, the efficiency is better. There is no
quick way to remove thrombin or GST. Any way if you want to have high quality
protein, you have to use chromatography or HPLC. In my case, after monoQ, I
got single band coomassie blue band on SDS-PAGE.
degraded
any
be
protein |
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r****o
发帖数: 105 | 16 BDscience有专门表达GFP fusion蛋白的质粒卖。比如pEGFP系列。
我用过pEGFP-N1,感觉不错。你不妨试一试。
另外，GFP其实是个结构蛮紧凑的蛋白，即使没有很长的linker，
也应该不会干扰你要表达蛋白的折叠。比如我用的pEGFP-N1的多克隆位点
实际上与GFP就没有专门的linker sequence.但是表达出来的fusion protein
活性很好，而GFP折叠也没有出问题，免疫荧光实验看得很清楚。
如果你实在想加一个linker sequence,可以参考一下pEGFP-N1 多克隆位点
序列。
另外刚才查了一下文献，感觉linker design还是很有讲究的，对于较短的
linker,原则是要用flexible 和hydrophilic氨基酸。Flexible 的氨基酸，比如
Gly和Ala; hydrophilic的氨基酸比如ser。
有些情况比较复杂，较短的linker不行。比如有报道表达一个protein G
和luciferase的fusion protein，中间的linker是GGGGS。发现虽然luci |
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j*******8
发帖数: 933 | 17 Ru-Ti.
I'm testing protein interaction in the post-synaptic density (PSD) fraction
of brain tissue. Usually, we use 1% SDS to dissolve the PSD pellet. However,
we're now planning to test protein interaction with IP. Can anyone tell if
1% SDS interupt protein interaction? what about 0.1%?
Thanks! |
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w******e
发帖数: 1187 | 18 希望知道某个protein的surface level绝对值（即×××copies）而不是相对值。
also BTW，有人知道total cell surface protein level的量级吗？ms通常每个
protein有10e4~10e6 copy，大概有多少种呢？
多谢～ |
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A******y
发帖数: 2041 | 19 Recombinant protein is around 300 a pop...if you need a lot of protein, you
need to make it yourself. Also, does activity matter? If you are doing
structure, making protein should be a piece of cake or you are in the wrong
lab. Or you can call the company for a larger scale quote. I used BPS biosciences, and the owner will do his best to make a sell and will try to make thing works, but I bet it will still cost you.
or try addgene and hope the plasmid of interest is deposited and make it
you |
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w******e
发帖数: 1187 | 20 covalent linked by EDC/NHS。想用一种universal的方法（非ELISA）测
beads上link的protein amount。因为beads本身会有interference，想要一种
assay方法，能根据protein amount给出signal，but the signal can't be
together w/ protein, so that the beads can be removed b4 measurement.
any thoughts? Thanks in advance! |
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w******e
发帖数: 1187 | 21 要把target protein conjugate到beads上用于下游的separation。现在在试
dynabeads，发现conjugation efficiency远低于预期值。而且比较头疼的是
要测出真实efficiency也很tricky。间接测反应后溶液里剩多少protein很
不准。
请高手指教有没有efficiency比较高的beads？最好是magnetic的（是不是
只有dynabeads系列？？），其他的介质也可以。最好是covalent linkage，
biotin-avidin的也行。BTW我的target protein pI在5~6
非常感谢。 |
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K*C
发帖数: 2825 | 22 我最近做了一些fusion proteins.我的建议是你不要在引物里面加linker。
最好先做出一个Target protein-non coding region-Fluorescent protein（FP）的
prototype来，然后用Site-directed mutagenesis往里加linker。 FP加在N端还是C端
，是要试的。linker用多长的，什么样的，也是要选的。否则会对蛋白折叠有影响。 |
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w******e
发帖数: 1187 | 23 用radiolabeled RNA aptamer+varying amount of protein，gel shift测affinity，
develop出来所有的protein concentration都只有一条很靠近加样口的band（很
怀疑根本没进gel），而且确定跟aptamer alone的band不是一条。请教有人遇到
这种情况没有？and如果确定了那条band根本就是在加样口附近，问题怎么解决？
protein确实挺大（250kd），但不应该进不了gel。
//bow |
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s******9
发帖数: 283 | 24 着急寻找三种protein complexes，hetero-oligomer或者homo-oligomer无所谓，
protein最好要小，能在E.coli体系表达。希望三个complexes Kd值分别在pM，nM和uM
的级别。 |
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p**u
发帖数: 138 | 25 I know a friend who met similar problem (tried strain for codon bias, toxic
protein, none of them worked). He deleted about 10 amino acid at the N-
terminus of the protein, then the protein was expressed well in BL21 (DE3). |
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s********n
发帖数: 2939 | 26 Then I moved to culture more and lysed the cells using a sonicator, after
resuspending the pellet with 1x cold PBS/1% PI. After all these, I loaded
the supernatant and pellet side by side and stained by GelCode and a bulk of
protein (I assume it was protein) was detected in the pellet part.
你有没有在supernatant中检测到你的target protein？如何检测的？WB or activity?
从你的表述你好像没有做WB。
Then I tried 1L culture and repeated the 2nd experiment. This time, I
incubated the supernatant with glutathione sepharose 4b beads an... 阅读全帖 |
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x*******i
发帖数: 32 | 27 Hello There,
We have a protein (pI = 8.9, 10 mg/mL, MW 100 K) precipitated in 10 mM
phosphate buffer (pH around 7) after overnight storage at 4 degree. The
precipitate will be soluble again when warming up solution to room
temperature. Addition of NaCl will prevent precipitation. Other common
buffers are OK not causing precipitate.
Hypothesis is ionic cross-linking. Phosphate ion has intense negative charge
density and working as the cross-linking agent, its multiple negative
charges interact wi... 阅读全帖 |
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m**e
发帖数: 857 | 28 can you believe of it? Just 3 amino acid.
Regulatory evolution through divergence of a phosphoswitch in the
transcription factor CEBPB
Lynch VJ, May G, Wagner GP.
Nature. 2011 Nov 13;480(7377):383-6. doi: 10.1038/nature10595.
Abstract
There is an emerging consensus that gene regulation evolves through changes
in cis-regulatory elements and transcription factors. Although it is clear
how nucleotide substitutions in cis-regulatory elements affect gene
expression, it is not clear how amino-acid sub... 阅读全帖 |
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c********r
发帖数: 189 | 29 1) FRAP to see the protein's mobility in nucleus.
2) cell fractionation to see if the protein only in DNA binding fraction |
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l**********1
发帖数: 5204 | 30 plus if your nuclaer protein is Proline-Rich Homeodomain protein (PRH)
you can try oligomeric assembly
Please go to 2010 one paper:
//www.ncbi.nlm.nih.gov/pubmed/20675722 |
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l**********1
发帖数: 5204 | 31 RE: 德艺双馨的楼主
please go to
//www.creativebiomart.net/description_10243_28.htm
You Position: Home >> Antibody >> Polyclonal Antibody >> Rabbit PAb >> Anti-Cdh5 PAb
Anti-Cdh5 PAb
Cat. No.
CPB-133RM
>
Immunogen
Recombinant mouse CDH5 protein.
Antibody Type
Rabbit Polyclonal Antibody.
Ig Type
Rabbit IgG.
Formulation
0.2 μm filtered solution in PBS with 5% trehalose
Preparation
Produced in rabbits immunized with purified, human cell-derived, recombinant mouse CDH5 extracellula... 阅读全帖 |
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t******t
发帖数: 36 | 32 Is there other suggestion about study protein-protein interaction? Thanks |
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f***o
发帖数: 88 | 33 google NaCl protein-protein interaction
this way can help you learn specific knowledge quickly |
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l**********1
发帖数: 5204 | 34 try refer
Next Generation Wet lab protein-protein tools review:
//www.ncbi.nlm.nih.gov/pubmed/21999828 |
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e****e
发帖数: 1042 | 35 就大小不一样呗！我要的产物是3kD左右的peptide，其它的都是杂质，我想知道比如大
于10kD的protein的量是多少，就算是protein的杂质量。我不需要分离它们。 |
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e****e
发帖数: 1042 | 36 As I said before, I've worked on protein projects for a long time. But I
never think too much. Once a person who never works on this field ask such
an question, I lost for a moment. Just like when a kid saw a piece of white
paper turns to black when the paper was burned, he asked why the white turns
to the black. I told the kid the fire make the paper to carbon. Then the
kid said: "So the paper is not carbon?" I suddenly realized the paper is
also made of wood and it should be carbon. Then I nee... 阅读全帖 |
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b******y
发帖数: 627 | 37 If you have a 50 a.a peptide with no particular 2nd structure, I call it a
peptide. Some polypeptides, like ww domains, have only about 40 a.a. (3
short beta strands packed in an antiparallel fashion), which can bind
proline-rich peptide ligands. I can call them proteins or at least protein
domains. I believe there are a few domains that are of comparable size or
even smaller than ww domains. Don't remember what they are called on top of
my head.
As we all know, it is so trivial to translate DNA... 阅读全帖 |
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b******y
发帖数: 627 | 38 Finally to LZ, you can quantify your product using HPLC at any stage of your
purification. Meanwhile measuring total protein including protein, peptide
and your product using branford or methods alike, you will know the
enrichment or purity at any stage. I believe this is more or less how your
company is doing it.
What makes me agitated is:
Even if you can magically distinguish 3 KDa peptides from 1-2.9 KDa and 3.1
KDa-infinite, you still don't know whether they are purely from your product
or p... 阅读全帖 |
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p***g
发帖数: 66 | 39 我从文献中得到了一些疾病相关的SNP(single point mutations, for example,
rs85462)和对应Gene. 我想看看这些SNP都是什么类型: frameshift, missense,
nonsense etc. 该查什么database, db.snp?
然后我想作图看看mutations 分布在哪些protein domain (象附件图). 需要用什么
databases (protein domain mapping) 和什么软件. 请各位多多指点, 万分感激!
A little more information: I need to check several hundreds of SNPs for tens
of genes. Just visualize their distribution and effect on protein function/
structure. That’s why I need some software/web applications to do it. |
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c*********r
发帖数: 1046 | 40 哪位TX知道怎么样处理protein A resin 可以尽可能减少下一次样品的交叉污染。
protein A resin 比较贵， 要反复用， 但是最近发现提取的蛋白不纯， 混有上次的
样品。 怎样REGENERATION 才能 减少污染呢？
谢谢！ |
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f*******0
发帖数: 62 | 41 【 以下文字转载自 Faculty 讨论区 】
发信人: fishman80 (最爱鱼儿), 信区: Faculty
标 题: 请教几个用来产生 membrane proteins设备的价格
发信站: BBS 未名空间站 (Mon Feb 25 13:44:06 2013, 美东)
有幸得到一个AP position， 现需要估算个 startup package。 因为我主要是用E.
Coli express and purify membrane proteins，特向有经验前辈们请教几个主要用到
的仪器大概价格 （有个区间就行）。多谢多谢！
1. High performance liquid chromatography
2. 1L bacteria incubation shaker
3. OD spectrometer (for bacteria concentration measurement)
4. Allegra type centrifuge (for protein solution centrifuge／concentrate)
5. Bacteria ce... 阅读全帖 |
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i*S
发帖数: 175 | 42 RNA --- in situ hybridization
Protein --- 免疫荧光
除了这两个，就没什么可以用的了吗？
现在我是做免疫荧光发现某个protein存在于某神经组织的一种细胞里，但是以前都认
为是只存在于这个组织另一种细胞里的，老板不放心想确认一下.这两种细胞也是离得很
近的，很难分离以后单独检测RNA/protein什么的。我觉得是不是做个in situ看看mRNA
以外就没什么办法了啊？ |
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s******s
发帖数: 55 | 43 请教有什么方法可以concentrate 低浓度的protein sample, 并且有较高的recovery
rate?
我的protein sample是cell lysate, 浓度大概是40ug/ml,总共3ml左右.想concentrate
到2mg/ml左右。试了millipore 的Amicon column, 是可以concentrate到想要的浓度，
但是回收率只有60%左右。大概很多protein粘在了膜上。因为最后的蛋白量对我很关键
，请问有没有什么方法可以达到80-90%的回收率？
十分感谢！ |
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s********n
发帖数: 2939 | 44 需要做一些high throughput protein identification, 刚接触这个领域，除了mass
spec外，是不是就是protein microarray？还有没有其他一些技术？
对protein microarray不了解，有没有这个方面公司的产品可以推荐？万望邻域专家不
吝赐教啊。 |
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s***y
发帖数: 24 | 46 【 以下文字转载自 Immigration 讨论区 】
发信人: sevny (forever), 信区: Immigration
标 题: guest editor 机会，protein interaction
发信站: BBS 未名空间站 (Thu Aug 28 21:32:49 2014, 美东)
guest editor机会.
Journal: Biochemistry insights journal supplement (Libertas Academica)
Scope: Protein binding and protein interactions
Tasks: 1. Solicit five papers
2. Compose an editorial
如果有兴趣，请站内联系。 |
|
A*****e
发帖数: 29 | 47 You need a database to exclude non-specific binding proteins. What epitope
did you tag on your protein? According to my experiences, more than 95%
proteins are not your interest. Your may try CRAPome online program to give
you a clue what are real hits. |
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j*********j
发帖数: 124 | 48 不知道你到底要做什么。。。
我猜，你是不是先要用protein/nucleosome-protein docking 找到binding interface
， 然后才能估计binding affinity啊？ |
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M******g
发帖数: 152 | 49 Try not to heat up your protein samples in SDS loading buffer. Many
membrane proteins form aggregates even in SDS-loading buffer when they are
heated. 70 kd may be your protein aggregation. |
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M******g
发帖数: 152 | 50 Try not to heat up your protein samples in SDS loading buffer. Many
membrane proteins form aggregates even in SDS-loading buffer when they are
heated. 70 kd may be your protein aggregation. |
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