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Upregulating Aquaporin-1 in Bovine Aortic Endothelial Cells with Arginine - scr


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#1 cbraval

cbraval

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Posted 12 March 2009 - 09:45 AM

Hi I need a little help in designing an experiment. I am working with bovine aortic endothelial cells and I want to upregulate Aquaporin-1. I found a paper that showed aquaporin-1 was upregulated in first trimester-derived extravillous cytrophoblasts with arginine vasopressin (AVP), a hormone. At a concentration of 0.1 nM AVP at 10hrs, a 4 fold protein expression increase was reported by means of a western blot and PCR. I want to utilize AVP in my BAEC cells. I corresponded with the author and he said the effect likely will be different since I am using a different cell line. However my advisor seems to believe it would be worth trying and hopefully we see some upregulation.

I am using 12 well plates, and similar media formulation (Minimum essential media, fetal bovine serum, penicillin streptomycin, L-glutamine). I have 1mg of AVP, which has a MW of 1084.23. I plan on diluting all of it in 500mL of double ionized water and then aliquot enough into 50mL vials of my media to obtain desired concentrations. I was hoping to get a little feedback on my design. I think if I do 0.1nM, 1.0nM, and 10nM I would have a good starting point to see the effect over this range of concentrations. Any feedback would be appreciated.




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