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Hope you managed to get some via the meeting this week. Let me know if you need me to do anything.

Re: Cell Line request by HayleyWHayleyW, 04 Mar 2010 10:55

Does anyone have an aliquot of either VCaP or MDA PCa 2b cell lines I could have and do you know if they express endothelin 1 receptor?

Cell Line request by Mickbrown1Mickbrown1, 01 Dec 2009 11:01
Cell Lines
Mickbrown1Mickbrown1 01 Dec 2009 10:57
in discussion Methodology / Anything else » Cell Lines

Does anyone have an aliquot of either VCaP or MDA PCa 2b cell lines, derived from bone mets, that I could have. Also do you know if they express the endothelin-1 receptor?

Cell Lines by Mickbrown1Mickbrown1, 01 Dec 2009 10:57

I doubt that a polytron would break up cells, unless you were using it with a strongly chaotropic buffer. However, i'm not sure i'd use one if my goal was to make high MW genomic DNA. I would guess that it is no more 'aggressive' than using a dounce, but it is very good for mashing up such a tough and fibrous tissue, especially for RNA.

Re: Western Blot from tissue by AxelATAxelAT, 12 Nov 2008 20:54

Will the polytron ruin the DNA/chromatin and protein complexes? How aggressive is it? I've never tried it so I was just wondering.

Re: Western Blot from tissue by HayleyWHayleyW, 11 Nov 2008 19:28

In my experience you need a polytron to realy mash up prostate; it is surprisingly tough and fibrous. I would recommend using a polytron and homogenising in sample buffer, or use trisol (to make RNA) and 'recover' the protein fraction (i've never tried this and suspect it is not as good). The downside is that it's hard to measure protein concentration if you make your extract in SB…

Re: Western Blot from tissue by AxelATAxelAT, 04 Nov 2008 09:40

I need to get samples for WB from human TURPs. Which is the best method to get these prostate samples homogenised? I thought of freezing them with liquid nitrogen and then use a mortar to convert it to dust and then lisate it. The douncer did not really help.

Western Blot from tissue by Antonio R-MAntonio R-M, 31 Oct 2008 18:15

Does anyone have a urine collection with patient outcomes?

I have an ELISA which works well in urine. Our current dataset is too young to have any outcome data so I would like to be able to test my hypothesis.

Even a very small cohort would be very useful.

Urine samples by HayleyWHayleyW, 30 Oct 2008 11:26

Could it be that there is a better alternative to arrays ?

http://www.nature.com/news/2008/081015/full/455847a.html

Well, since arrays are essentially non-radioactive-reverse-dot-blots this would not be very surprising.

see also

http://www.nature.com/nbt/journal/v25/n8/full/nbt0807-878.html

the death of arrays ? by AxelATAxelAT, 17 Oct 2008 14:33
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