TLDW: 1. put sucrose in a tube, with low density at top, high density at bottom. 2. add some proteins (with markers). 3. spin really fast. 4. now less dense proteins are at top, more dense at the bottom.
The difference is: the CsCl gradient (solvent) and the proteins gradient is created at the same time, I mean, you mix and distribute evenly both of the solutions; after that, you apply centrifugation. On this other method using sucrose, you have to create the gradient of solvent (sucrose) by centrifugation, and after that, you can apply the molecule you want to separate by density (you apply it like a coat of protein suspension on the top of the sucrose gradient).
TLDW: 1. put sucrose in a tube, with low density at top, high density at bottom. 2. add some proteins (with markers). 3. spin really fast. 4. now less dense proteins are at top, more dense at the bottom.
Gained immense pleasure watching you struggle to spell sucrose. Liked, subscribed, shared.
You are saving my degree. Thankyou!
The explanation was excellent.
Very informative. thank you
Very helpful thank u soooo much
Awsome video that remove all my ambiguities
Thank you very much...
This was very helpful, thanks for making this video :)
You saved me, thank a lot !!!!!
good lecture
Is the sucrose gradient high-speed centrifugal column commercially available?
Yes, there is a company called Alfa wassermann that makes them.
thank u!!!!!!!
Great video!
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It's perfect video!
I have a question
what is different between this centrifugation and CsCl2 density-gradient centrifugation
?
The difference is: the CsCl gradient (solvent) and the proteins gradient is created at the same time, I mean, you mix and distribute evenly both of the solutions; after that, you apply centrifugation. On this other method using sucrose, you have to create the gradient of solvent (sucrose) by centrifugation, and after that, you can apply the molecule you want to separate by density (you apply it like a coat of protein suspension on the top of the sucrose gradient).
Why sucrose is used???
It's non toxic, cheap and probably non-polar.
Also, sucrose molecules probably don't diffuse as fast as salt molecules, due to their larger size.
Good info❤️
I thought you were going to show the mechanics of actually making the gradient. Disappointing.
You just centrifuge the sugar solution for a couple of days or so
Just spin it