Greetings and all for my first post here!!

A little back story. I am working with HEK293 cells and HERG channels. Bright and early one Monday morning I went to retrive my pipette solution from frozen storage and the freezer had quit working. The thermometer was reading warmer than room temp, and computer records showed that the freezer had gone out sometime on Friday morning. So I know that all my pipette solutions are trashed.

Since the MgATP is the main temperature sensitive component. I have entered search queries in PubMed with MgATP and thermal stablitiy. But I get a ton of articles that deal nothing with pipette solution. Is there any articles that might deal with ATP from other sources in a different use from pipette solution? So if someone can point me in a direction of an article which deals with the time that pipette solution (or MgATP in general) can remain at room temp before becoming completely trashed, that would be greatly appreciated.

Thanks!!

Id say toss it. MgATP is good for 1 week at 0C according to the Sigma product sheet, so I doubt it lasted the weekend.

Sorry

Rb

I keep my MgATP containing solutions on ice for no more than 6h while I am recording...then I consider them useless. Aliquot lots of vials of pipette soln in small volumes and maybe get a stratacooler or similar cryostorage box to put in your freezer that will thaw much more slowly if your freezer breaks down again.

Thanks for the responses. I agree the solution is trashed. But didn't know if anyone had read a paper or had heard of a "fish tale" of someone using MgATP past a day. I think we have all heard someone say something that would rasie an eyebrow. :P

I tend to keep the pipette solution for no longer than an eight hour day. In the past I have kept it on ice and at RT with little difference in the performance of the experiment. However, I like the idea of a cooler box, but more for keeping the syringe on the bench and as cool as possible without freezing.

To prevent future down time I am going to split the solutions into three groups and store in three different freezers. I knew mom was right when she talked about all the eggs in one basket. Moms are so wise!! :)

Thanks again guys!! Back to lurking!!

If the freezer (-20C) works well, how long can we keep MgATP containing pipette solution in general? 1 month? 3 months? 6 months? Or even more?
Thanks.

First sorry for the threadomancy. I've been swamped with stuff and not able to hang out as much as I would like.

Golden Ball, the pipette/intercellular solutions I make I keep for a month and then trash.

For those that are interested. I rigged a couple of frozen gel packs up to hold my syringe during the course of the day and keep the pipette solution nice and cold (read: almost frozen) and the recordings I got were outstanding! Fraser, thanks for the idea about the stratacooler, which spawned the idea of the gel packs....