My shittiest method: The in vivo complex of enzyme (ICE) assay.

My shittiest method: The in vivo complex of enzyme (ICE) assay.

In this new challenge reseachers/science students have to present the worst method they are forced to work with.

The Challenge

My fellow lab-rats!
While I am sure that you love working in the lab every single day of your existence (that’s sarcasm), it’s a sad fact that we all suffer from having to conduct experiments that are exhausting, time-consuming and fail to work spontaneously.
Take this challenge as a possibility to rant about your worst assay, get at least some fun and rewards our of it and maybe win the sympathy of the STEM world by being declared the poorest lab soul of all!

The following rules apply:

  • describe the worst method/assay/experiment you have to work with
  • use the #myshittiestmethod and the #steemstem tag
  • use “My shittiest assay method: … ” as title
  • you can nominate some peers, they have to participate then (unless they want to be seen as very, very lame). But you can join the party without invitation aswell!
You can win:
  • a 100% chance to get my (albeit almost worthless) instant upvote!
  • what I deem the shittiest assay by one month from now wins 200 SP as delegation... or as much delegated SP as needed to reach 500 SP (which grants the voting power slider) for as long as you need to reach the 500 yourself (yes, this means people with more than 500 SP already are participating just for the fun – but you can keep the rewards too^^).
I’m starting today, and I nominate @alexs1320 – I’m quite sure microscopy isn’t just fun and taking pretty pictures! (And from his anti-vaxxer posts, he seems able to do a quality rant!)^^
And there it is - my shittiest method:

The ICE assay – why is this still a thing?

If you want to understand why we have to endure something like the ICE assay, you should first recap my recent post on topoisomerases (Topos).
As I wrote there, Topos are incredibly important enzymes for regulating DNA topology, and if they are inhibited this can cause terrible consequences.
Now, measuring the inhibition of Topos in a cell-free experiment is quite easy – you just buy the enzyme, incubate it with a suspected inhibitor and some supercoiled or katenated DNA, then you separate the DNA by electrophoresis and you see whether the enzyme still works or not.
Ofc, it’s trickier than it sounds, but a lot easier than monitoring the situation in living cells or even animals. Also, there are two types of inhibition: the “catalytic” inhibition and Topo poisoning.
And the only way that I am aware of to detect Topo poisoning in vivo is to detect the complexes of Topo and DNA that are accumulating when a Topo poison is present. And that’s when the ICE assay falls on a researcher’s head like the very sword of Damocles.

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