i want to use 3 MA for stopping autophagy in my N2A transfected cell line overexpressing Parkin....but the 3MA that i bought shows on it 10 MIlimole/militre in DMSO? so how can i use it?any idea please?
There is a publication (http://www.ncbi.nlm.nih.gov/pubmed/24710924) in which the authors used 10 mM 3-MA to inhibit autophagy in N2A cells. But I think that this is the final concentration. According to the Sigma Aldrich website 5 mM is most commonly used. So you could try to work with 5 mM. Your 10 mM stock solution is not concentrated high enough to apply a 10 mM final concentration. Better to use the crystallized product and dissolve it in DMSO according to your needs (cat. no M9281 (Sigma-Aldrich), for example).
Regards, Christian
You can dssolve the powder directly in your culture medium to achieve 10mM. It will not dissolve easely so you want to make sure that your medium is at 37C and vortexing a couple of times will speed things up. Then you can either use this medium directly on the cells (this is how I do it) or you can dilute it 1:1 and work with 5mM.
well thanks ever so much but my problem is that 3MA that i got is not powder,its liquid and says on its bottle 10mM/ml in DMSO and its altogether only one militre? somebody told be its concentrated solution of 1ml that contains 1000 microlitre?
well if it is already dissolved in DMSO at 10mM then you cannot use it for autophagy research on cells. The maximum DMSO concentration you can use on cells is 0.1% (1 uL per mL) which would give you a concentration of 1 uM and that is just way too low for autophagy research.
I am afraid you will need the powder..
We usually dissolve 3-MA directly in the media at 5 mM
Good luck with your experiments!!
Carlos
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