Centrifugal ultrafiltration (spin concentrators) for protein (or nucleic acid) concentrating

[@SouthAfricanCannabisExtraction]

Honestly amazed how you keep coming up with your topics. Wish this channel was around when I studied Biochem. Awesome resource! Keep up the great content!

[@danielreis3846]

Greetings from Brazil. Great video. I'm going to perform this centrifugation for the first time and your video helped me a lot.

[@thebumblingbiochemist]

Thanks! Glad to hear it!

[@eruvee]

This was great, thank you so much! Helped me a lot when I was asked to use them in a protocol for the first time!

[@thebumblingbiochemist]

So happy to hear - thanks!

[@user-hd5he4cf3n]

10k watching, thanks for your introduction🦋,love from China

[@thebumblingbiochemist]

Thanks! Happy to help. Good luck!

[@tonimaakter6673]

i love your videoas and explaination, they are really helpful always for my lab work😍

[@thebumblingbiochemist]

So glad they're helpful!

[@originalAtreyu]

have to mix them often. there gets to be a large concentration gradient some proteins will begin to crash (precipitate)
👍

[@SeriousBartYT]

Thanks for sharing another useful video. I usually go through some of your content before doing "simple" stuff like Transformation or Purification.
One small question : Lets say that I am using a Centrifugal concentrator with a 10kD cut-off. After adding the protein-containing solution to the tube, do you know of a method to estimate the optimal speed at which the protein in the tube would get concentrated in the least possible time ?
P.S. Based on some of my internships, I learned that the estimating the optimal speed at which a protein can get concentrated is usually determined by a trial and error basis. But I was curious if there was something I was missing.
- Bart

[@thebumblingbiochemist]

Glad you found it helpful! The manufacturesrs' website should have guides for that I think. Good luck!

[@darklight9827]

Thank you so much!

[@shuangshuangyang3043]

Hi! Thank you so much for sharing. It's super helpful. I'm wondering what speed you use for the swinging-bucket centrifuge?

[@thebumblingbiochemist]

Great to hear it was helpful! You can go up to 4000 g with these

[@roxy1963]

Thanks!

[@phumlileprettymamba5509]

Very much helpful. May you please assist, i'm need to separate laccase, manganese peroxidase and lignin peroxidase from crude extract based on size exclusion. Which column and buffers would be ideal for this experiment?

[@thebumblingbiochemist]

Glad you found it helpful! I don't have experience with those specific things so I recommend you try to find some published protocols for it. Sorry!

[@MadalenaGrenhas]

Hi! Thank you for all the tips and info about these devices! When reusing the Amicon 15mL (that looks like a 50mL falcon), what volume of a 20% ethanol solution should I add? 5mL is enough? Also, at what temperature should I store these devices between uses? Thanks

[@thebumblingbiochemist]

I'd just make sure there's enough liquid to keep it wet. And I kept in fridge.

[@HoaHoa-bf8gk]

Hi! Thank you so much for sharing. I have a problem at this step in my experiment. I need collect MWs

[@thebumblingbiochemist]

I don't understand what you're asking sorry but good luck!

[@HoaHoa-bf8gk]

That mean, ~200mg protein hydrolysate. What volume of 20% Ethanol I should add? From 10mL enough?
I don’t know to do centrifugal ultrafiltration that is there a concentration for sample solution?

[@rogsang9764]

thanks :D

[@davidreyes346]

Hello, with what solvent should I wash the filters to reuse them?

[@thebumblingbiochemist]

I think I used to use 20% EtOH