Biochemistry's topics - tribe.net

HEY YOU, yes u, why don´t u post...... POST PLEASE ;-((((((((((((

Prall — Fri, 27 Jan 2006 15:18:03 GMT

Dear all members. I wonder why noone likes to answer my posts. Any excuses, any explanations???
Please answer this post..............PLEASE ;-(((

posted in Biochemistry - 5 replies

cDNA clones, buy them cheap, not like the Origene clones...

Prall — Wed, 30 May 2007 18:43:34 GMT

Hey Guys,
I found a very good plasmid database! One can buy cDNA clones, shRNA , genomic fragments
and other transcripts very cheap, It´s PlasmID, plasmid.med.harvard.edu/PLASMID

Check this out !!

Best regards,

Seb

posted in Biochemistry - 0 replies

looking for cDNA clones from receptor tyrosine kinases

Prall — Sun, 18 Mar 2007 14:27:21 GMT

hey lab folks,

i need cDNA coding sequences for some receptor tyrosine kinases, here they are:

ALK
FLT4
RYK

if you can help me, please answer or contact me directly.

don´t mind for the costs, i´ll pay the shipping and taxes!!!

thanks in advance,

ron

posted in Biochemistry - 1 reply

Vitamin D question

Grandma — Sat, 16 Dec 2006 07:32:55 GMT

Is anyone familiar with the tests required to determine circulating 25(OH)D levels to determine someone's vitamin D nutritional status? Price, how long the results take, what labs to use, etc?

Thanks,
Grandma

posted in Biochemistry - 0 replies

Serious question for the lab folks...

Dr. Dave — Thu, 16 Nov 2006 01:10:36 GMT

Can anyone recommend a good vendor(s) for some analytical instruments?

I need a conductivity probe, 0 to 2000 uS/cm
thermocouples, -5 to 35 C
and a flow meter that can measure as low as 20 ml/min but can be scaled up.
They all have to communicate with some type of Data Module with a USB port.

Not a joke. I need them for a project and I haven't done this sort of thing in years.
I am going crazy sifting thru vendor webpages and am looking for some shortcuts.

Dave

posted in Biochemistry - 4 replies

cancer research in industry-- I'm looking for a position..

brent — Mon, 03 Jul 2006 22:33:00 GMT

hi all, not to "spam" too much but I will probably post this elsewhere as well, but the bottom line is I'm trying hard to get into an industrial research position in cancer. I have a ph.d. in cancer biology (from univerisity of Arizona) , and have done 1 postdoc at the university of colorado health sci center. the postdoc was in neurodegeneration, but focusing on apoptosis which is what I'd say my 'specialty" is; so even though I didnt' stay in cancer I studied similar signalling pathways as I did for cancer. I wanted to get back into cancer research, though, so after ~2 yrs of the postdoc I became a research associate in a lab that was doing breast cancer research (still at UCHSC). unfortunately that lab moved to TX and also I just came to realize I really didn't want to stay in academia.

I've had four on site interviews now: Centocor (part of J&J in PA), Telik (san fran), Astra Zeneca (boston) and Southern Research Institue (AL). I know its not entirely this, but I feel like part of the reason I make the short list of on site but then dont get the hire is lack of industrial experience.. so I'm in the catch 22 of needing industrial experience but no one will hire me to GET said experience. I'm not blaming "them".. I realize, too, unfortunately, that I'm not daily immersed in cancer research anymore (I'm currently working at a small biotech doing protein purification; something I chose to do while I look for a cancer job so as to expand my skill set) and so I feel like the longer I'm away from cancer research I'll probably not be as "sharp" on interviews. I try to keep up but its hard being on "the outside" and reading journals; its a lot different than being daily immersed in thinking about cancer.

sorry to blab. I hope if someone reads this and is working in industry (specifically cancer research) they might contact me so I can get my resume to them, and any tips on how to 'break in" to an industry job, post-ph.d.... (I have applied sporadically to industry post-docs and am looking at them more seriously but this far haven't gotten an interview for one, and frankly would prefer to not have to do another postdoc, but its starting to be where my overwhelming motivation is to get back into cancer research, whatever my "title" is...

thanks so much for any thoughts, feedback, networking, etc,
Brent
bdb777 at hotmail dot com

posted in Biochemistry - 0 replies

For what reason are u interested in Biochemistry?

Prall — Thu, 02 Jun 2005 22:34:04 GMT

Everyone joining this Tribe should give a SMALL overwiev what he is doing or thinking around the theme Biochemistry.
Do u work in science/industry? If yes were? What are u doing there? and so on.
So who likes to start??

posted in Biochemistry - 12 replies

Biochemistry, future purposes and interesting subject/

Prall — Wed, 15 Mar 2006 01:03:48 GMT

What do you think Biochemistry could do for the manhood?
Are you interested in the future investigations of biochemistry? Why?
As the whole "cloning" theme semms to be a controverse discussed theme:
What do you think of cloning creatures (plants or animals)
What do you think is cloning and what are the (dis)advantages of cloning compared to classical plant/animal breeding ?

posted in Biochemistry - 1 reply

What do/did u do in our daily research?

Prall — Fri, 20 Jan 2006 21:01:12 GMT

I investigated the three NLS (Nuclear Localisation Signals) of the human ribosomal protein S6. I tried to elucidate either all three NLS are necessary to ensure nuclecytoplasmatic transport or if one or two NLS suffice.

In summarisation it can be said that more NLS lead to higher nuclear import effectiveness.
The degradation of ribosomal proteins in the cytoplasm requires a high-speed nuclear import.
One can assume that several NLS in one Protein increase the rate of nuclear import. This was shown in my diploma thesis
Additionally several NLS in one Protein could help to target the nucleus by different transport receptors, which means different Importins.

For that I designed mutants of this rib. protein that contain other amino acids in the place of the NLS1 or NLS2. People of my working group already designed other mutants, so that now ALL permutations of mutated NLS are present.
I use DNA-Constructs, which encode for EGFP-S6-Fusion proteins, that means EGFP is coupled to the N-terminal of the human ribosomal protein S6. DNA-Constructs that encoded for two NLS were designed and cloned by myself in my diploma thesis at Institute for molecular cell biology II, University Hospital Hamburg, Germany.

The DNA of the Constructs was transcribed "in vitro" and the isolated mRNAs were then "in vitro" translated. There for rabbit reticulocyte lysate was used. The synthesized EGFP-S6-Fusion proteins contained either no (negative control) one, two or three (positive control) NLS.

To investigate the potential of all EGFP-S6-Fusion proteins to enter the cell nucleus, an "in vitro" nuclear import system was used:
After Digitonin-permeabilisation of HeLa-cells, the Cytoplasma is washed out and is reconstituted with reticulocyte Lysate that contains the translated EGFP-S6-Fusion proteins. The cells were grown on cover slips. After the nuclear import system procedure they were mounted with Mowiol and the localisation of the EGFP tagged S6 fusion proteins in the cell was detected by fluorescence microscopy

Here are the results:
The positive control with three NLS showed definite coloured Nucleoli (Nucleoli are compartments within the nucleus!!).
The negative control with no NLS showed no definite coloured Nucleoli, but cytosolic coloration.
EGFP-S6-Fusion proteins that contain two NLS are imported in the Nucleus more efficient, than only with NLS 3.
The EGFP-S6-Fusion proteins with only NLS1 or NLS2 are not imported into the nucleus in vitro. Prior Investigations showed that EGFP-S6-Fusion proteins with only NLS1 or NLS2 are imported in vivo.

My conclusions can be used to investigate the different transport receptors that are involved in the nuclear import of the human ribosomal protein S6. Furthermore it can be investigated which transport receptor targets which NLS of the human ribosomal protein S6. Therefor the above mentioned "in vitro" nuclear import system can be used.

Greetings Seb

posted in Biochemistry - 0 replies