Hypoxia - downregulated HIF-1A
Posted 15 April 2010 - 04:59 AM
I have been tryin to study the regulation og HIF-1A for my hypoxic experiments. I tried doing cobalt chloride dose response, hypoxia incubators and bubbling up culture media with Nitrogen, and still HIF-1A is not up regulated in my cells. My other protein of interest which is a hypoxia response protein is upregulated, so to confirm that the cells are undergoing hypoxia, HIF-1A the marker for hypoxia is not upregulated...
can anybody pleeeeeeeeeeeeeeease suggest why this is the case and ways to rectify the problem??? I have been stuck with this for the past 2 years!!!! yes 2 precious years of my PhD!!!!!!!!!!!11
Posted 01 June 2011 - 06:50 AM
A question, what do you look for? HIF mRNA or protein??? Because under hypoxia, the first thing that happens is an stabilization of HIF protein, not an induction of HIF transcription... Sorry about my English
Posted 03 June 2011 - 06:38 AM
I never trust anything that can't be doubted.