The Seahorse kits are so expensive. I was wondering if I could just buy the chemicals and use these in place of the kits. Specifically Oligomycin and 2-DG.
Hi Helen,
I agree that the Seahorse chemicals are indeed very expensive and it is just a way for "Seahorse" which is now Agilent Technologies to make extra money. I would boycott buying or using the so called Mito Stress Kits or Glycolysis kits and purchase the chemicals separately from other suppliers. I have always used Sigma for Oligomycin, Antimycin A, FCCP, Rotenone and 2-DG. Just be sure to dilute and store the chemicals appropriately, things like oligomycin, rotenone, uncouplers, and antimycin A will need to be frozen, I suggest doing this in small volume aliquots to prevent freeze thawing - this will maintain the quality of the respiratory inhibitors for longer. The good thing is that it will cost about £200 to buy in the chemicals from sigma to run a typical 'Mito Stress Test' and they will most likely last for years when handled and stored appropriately.
Another thing to consider for your experiments is a different uncoupler (if using), seahorse recommend FCCP I think but BAM15 is likely a better option, as it is suggested be more specific for mitochondrial membranes.
Good luck and all the best
Jonny
That's what I wanted to hear!!! Thanks Jonny.
Which Oligo do you buy? The Seahorse kit has a mix of Oligo A, B and C. So I thought this one would be the best.
http://www.sigmaaldrich.com/catalog/product/sigma/o4876?lang=en®ion=AU
Is this the one you buy?
Thank you very much guys! I was looking exactly for this information. Jonathan, could you explain if you see many differences comparing the two uncouplers?
Thanks again!
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