| IHC |
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| 08:55am 06/11/2009 |
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Hey all - I have a sorta odd question regarding IHC. In the past, I've always done IHC on tissue on slides. My new lab's protocol (on 40um sections) involves doing all the staining on tissue sections floating in a 24 well plate, with little mesh screens under them that you can lift out, the tissues comes with it, and change from washes to primary antibody, and so on. The tissue is then mounted AFTER development, allowed to dry in the the hood, and then dehydrated through alcohol to xylene. This afternoon we're developing with DAB, and I'm accustomed to doing a hematoxylin stain after DAB. Think I should do the hematoxylin in the 24 well plate like everything else? I think if I were to try it after we mount on slides but before we dry them, the tissue would just fall right off the slides, but I also doubt that I could do it after we dry the tissue... Any suggestions? |
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Read 7 - Post |
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| Partial alkaline hydrolysis of RNA |
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| 01:50pm 05/11/2009 |
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Hello again.
I'm currently working on in situ hybridizations of frozen tissue slices from mice brain. We have an established set of probes and the like. Some work like a charm. Some don't. The main problem seems to be, with those not working as they should, that they seem to be too big.
The ones which have between 600 and 800 bp work fine with those with 1.2 kb don't seem to be able to enter the cells we want them to (I have nice signals in the cerebellum but I'm not at all interested in the cerebellum).
While we're working for the sequences to come back so we can check for restriction enzymes to use in shortening (Method 1) we also thought to use alkaline hydrolysis. Unfortunately it turned out that we have no signal on the gels whatsoever after performing alkaline hydrolysis and since we used everything treated with depc, it's unlikely we just simply lost the RNA to RNases.
We (adviser and me) used a sodium carbonate buffer with 40 mM NaHCO3 and 60 mM Na2CO3 at pH 9 (5 min at 60 C) and we suppose that the concentration might have been too high.
I looked around and found different buffers and found about a dozen different versions. Has anyone ever done it this way and could give me a pointer or something?
Thanks very much in advance. |
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| called democracy for a reason |
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| 11:14am 04/11/2009 |
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mood:  determined
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Look, I'm for gay equality. I firmly believe that the number and gender of your sexual partners doesn't matter. (Many things about your relationship may matter, but number and gender ain't in there - and I do *not* have the time to discuss this tangent right now.)
And I wish that Maine had voted for gay marriage, and I hope there won't be backlash.
But.
It really disturbs me when I see people who are supposed to be on my side saying things like "you don't get to vote on my civil rights." Actually, they do. Free speech applies to all speech, even speech that we hate. Likewise, democracy means the population gets to vote about whatever it wants. Even when they're wrong. Even when they're really really really wrong and their votes are screwing up your lives. Because "democracy, except when it gives an answer we can't abide" is not democracy at all.
[EDITED TO ADD:
Dear people from elsewhere, hello. Enjoy flaming me. Flames will not change my opinion in the slightest.
If you'd like to read more of the basic arguments against democracy, may I recommend Unqualified Reservations? (NOT THE COMMENTS, JUST THE ARTICLES.)
Also, for the zillionth-and-first time, I AM FOR GAY MARRIAGE.]
[EDITED TO FURTHER ADD:
Also, if you came here just to tell me how horrible and wrong I am, I would like to ask you why. Surely you're not expecting to convince me. So, what is the point of insulting a random stranger on the internet?]
[EDITED TO ADD, AFTER LAB:
Many of you seem to be reading this as "I get to vote on your rights [and you don't get to vote on mine, because none of mine are currently contested]." Let me give a counterexample. I have serious problems with the whole concept of 'rights' in general, as I've said several times in the comments. But, in my opinion, if *anything* is a right, if the concept of 'a right' has any *meaning,* then one of the most inalienable rights is self-defense. And yet, most of you would probably vote that I don't have the right to have a gun. And I'd think you're wrong (and I might well break the law), but I'd never say that you're not allowed to vote on this.
Also, the thing about free speech was an analogy. I know full well that voting and speech are different.
Also, I can't know how I'd feel if I were gay. This is true. But it does not affect the truth or falsity of my arguments.] |
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Read 91 - Post |
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| Cost of protein biosynthesis? |
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| 03:36pm 03/11/2009 |
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Hi guys, I want to do a bioinformatics analysis that includes the "cost" of each protein in the genomes I'm working on. I came across an article abstract where they say they did it, but the paper is *STILL* not out after a few weeks of my finding it, so I don't know how they did it.
I'm imagining it might just be as simple as using a "price" list per aa and adding up each per protein. Does this sound like anything y'all know about?
Thanks! |
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Read 4 - Post |
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| Paper search! |
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| 04:05pm 03/11/2009 |
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Sorry to be a pain, but does anyone have access to these?
Donald A Molony, W Brian Reeves and Thomas E Andreoli, 1989, Na+:K+:2Cl- cotransport and the thick ascending limb, Kidney International (1989) 36, 418–426
Gerardo Gamba, and Peter A. Friedman, 2009, Thick ascending limb: the Na+:K+:2Cl− co-transporter, NKCC2, and the calcium-sensing receptor, CaSR, Pflügers Archiv European Journal of Physiology, Volume 458, Number 1, 61-76
Rainer Greger1 and Heino Velázquez, 1987 , The cortical thick ascending limb and early distal convoluted tubule in the urinary concentrating mechanism, Kidney International (1987) 31, 590–596
krazykid197 @ gmail.com
Thanks in advance! |
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| Microscopy geeks: Organelle labels |
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| 01:28pm 02/11/2009 |
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Hello all,
I was wondering if anyone knows of a good non-commercial resource for gathering information about fluorescent organelle labels for microscopy?
I want to start compiling an easy-to-use resource for our microscope users but don't want it biased to a particular company.
Any help would be much appreciated! |
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| Career Satisfaction Survey |
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| 09:57am 31/10/2009 |
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I am currently a senior in high school. At my school, students are required to conduct a year-long senior thesis project in which they research a topic of their choice. I chose to research the current status of women in science careers.
Part of my methodology is to distribute a short survey to men and women in science careers to determine their attitudes toward their field and their work. I am interested in collecting data from ![[info]](http://l-stat.livejournal.com/img/community.gif) _scientists_. If you're willing to help a high school senior out, I would appreciate it very much if you would complete this short survey.
( Survey behind the cut... )
If you wish for your answers to remain confidential, you may send your answers to me through email at bursuz@gmail.com.
Thank you! |
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Read 43 - Post |
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| Immunoassay papers |
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| 11:33am 29/10/2009 |
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mood:  cheerful
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I was browsing some journal databases for some articles on immunoassays, how they work, and how one goes about creating one for detecting substances. I have only found a couple that would be applicable and it seems a very long and involved process (on the order of half a year). The journals I have found so far that might be relevant are:
Microarray immunoassay for the detection of grapevine and tree fruit viruses
Journal of Virological Methods [J. Virol. Methods]. Vol. 160, no. 1-2, pp. 90-100. Sep 2009.
and
Development and comparison of three diagnostic immunoassay formats for the detection of azoxystrobin
Journal of Agricultural and Food Chemistry Volume 54, Issue 3, 8 February 2006, Pages 688-693
I have read through these but I am still a bit hazy on creating, say, an ELISA.
Does anyone else have any suggestions. I am still in undergrad but I would like to become familiar with immunoassay's as I believe it will be very helpful to me in the future for creating a diagnostic field test I have in mind. Any resources on this subject would be greatly appreciated and do not have to be strictly journals.
Thanks in advance.Alexzander
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Read 6 - Post |
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| no subject |
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| 01:14pm 28/10/2009 |
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Hey Guys!
Would appreciate to get the article from IEEE!
My email: igor dot zlot at gmail dot com
Thanks in Advance! |
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| Article Requests |
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| 03:35pm 26/10/2009 |
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Hey guys,
once again I need some article help. And I need it on somewhat short notice (rather me and my new advisor do since we need to construct new tests), that's why I didn't try article_request first. If anyone could I (we) would be more than grateful.
Lee JW, Kim WR, Sun W, Jung MW; Role of dentate gyrus in aligning internal spatial map to external landmark; Learn Mem. 2009 Aug 25;16(9):530-6. Print 2009.
Gilbert PE, Kesner RP; The role of the dorsal CA3 hippocampal subregion in spatial working memory and pattern separation; Behav Brain Res. 2006 Apr 25;169(1):142-9. Epub 2006 Feb 7.
Gilbert PE, Brushfield AM; The role of the CA3 hippocampal subregion in spatial memory: a process oriented behavioral assessment; Prog Neuropsychopharmacol Biol Psychiatry. 2009 Aug 1;33(5):774-81. Epub 2009 Apr 16
email: seisbach [at] gmail.com
Thanks so much in advance. |
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| no subject |
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| 01:51am 26/10/2009 |
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music: sometimes you can't make it on your own by u2
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this was during mostly the end of the yankees game when they were finishing advancing to the world series, and as they won, happy times
it hung there like a dream I wanted so...
and I thought I might not reach it,
and that hurt,
but I'm reaching tonight,
and happiness is with it,
dear God, please,
it's so close,
aww, it tastes so good when you care,
when you're close,
and it's all you want,
it holds happiness,
that will carry me on...
it's so nice tonight...
I'm almost singing, clapping,
saying goodbye to the devil that haunted me...
almost smiling,
and then it's closer and closer...
a revelation,
and I got it,
we are the champions... |
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Read 4 - Post |
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