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Oof

I am starting a new round of cricket-poking. This time, with palmitic acid. On Monday, I asked about the injection vehicle, and wound up mixing up some phosphate-buffered saline, which I used to prepare the radiolabel.

When I injected crickets on Tuesday, I wasn't able to recover an adequate quantity of the radiolabel. I chalked that up to not being adequately careful when evaporating out the carrier (ethanol) and working with a small volume.

Wednesday seemed to go fine, so at the end of the day I prepared a larger volume of the radiolabel - enough to last about a week.

When I came in to get underway this morning, I first checked the quality of the radiolabel by putting small amounts into scintillation vials and counting the radioactivity.

No good.

When I looked at the solution, it was clear* that some of the palmitate had come out of solution. Argdarnit. I sat around for a little while and tried to read about tricks to use to get palmitate into solution. Eventually, my boss showed up, which was a relief because he has big sleeves that hold lots and lots of tricks. We eventually settled on the idea of adding some bovine serum albumin to the solution. I prepared a concentrated BSA solution (30 mg/ml) and then added a smidge to the radiolabel. Eventually things started to look better and I was able to get underway with the day's cricket-poking.

I didn't leave until 9:15 pm. Now I am waiting for bread to finish baking, after eating dinner way too late in the evening. Tomorrow is going to be painfully long.


*okay, it was _not_ clear and that's the point, heh.

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( 2 remarks — Remark )
randomdreams
Oct. 10th, 2015 01:46 am (UTC)
I would never have thought of BSA.
So how does palmitate enter their metabolic cycle?
rebeccmeister
Oct. 10th, 2015 10:02 pm (UTC)
Me neither! But it needed to be something that wouldn't affect the metabolic pathways we're interested in. Apparently another person who was doing these injections was using ethanol, but that just sounded way too wrong to me because ethanol will have all kinds of other effects.

I actually still need to go and check how palmitate fits into the picture. It captures an aspect of lipid metabolism, but that's all I know at the moment. Too busy collecting data, I tell you!
( 2 remarks — Remark )

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