Meet the AKTA! Fast Protein Liquid Chromatography (extended version w/extra tips for frequent users)

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Күн бұрын

If you’ve seen my lab bench in the middle of a protein purification, you know I can make quite a mess, but when it comes to the protein itself, I’m a stickler for cleanliness. And my main go-to protein cleaning tool is protein chromatography, a way to separate proteins based on differences in their properties. That bench mess can wait while I bring my protein to our protein “laundromat” - a FPLC (Fast Protein Liquid Chromatography) machine called an AKTA. So what’s actually going on in an AKTA? See accompanying blog post for text and figures bit.ly/aktainaction
If you want the original shorter video: 06/30/21 video: • Meet the AKTA! (and do...

Пікірлер: 25

@sunburntsatan6475 Жыл бұрын

Love the overview! Currently doing a protein purification using an AKTA system for a rotation I'm doing so this has been super helpful. If you're reading this now, it means you have already successfully defended--Congratulations!!

@thebumblingbiochemist Жыл бұрын

Great to hear. Thank you and best of luck with your purificcations!

@sakitlaatigue 2 жыл бұрын

Thank you for the elaborate information on FPLC AKTA. Currently I'm using a much simpler FPLC which is AKTAprime plus. However, the principle is still the same and so many tips and tricks I've learnt from your video.

@thebumblingbiochemist 2 жыл бұрын

Happy I could help! Good luck with your purifications!

@nicholaskampa2993 2 жыл бұрын

Very good and informative! I'm still learning how to use the machine, but videos like this help me figure it out

@thebumblingbiochemist 2 жыл бұрын

So happy I could help! Have fun purifying!

@user-bs3en7rh6v Ай бұрын

Great explanation! Loved it, thank you!

@thebumblingbiochemist Ай бұрын

Glad it was helpful!

@halilojaallways7204 Жыл бұрын

You Rock!! Very useful Information

@thebumblingbiochemist Жыл бұрын

Thank you! So glad it was helpful!

@JackieLiu0406 11 ай бұрын

Not one, not two...but four AKTA. I am amazed. 👍

@thebumblingbiochemist 11 ай бұрын

HHMI structural biology lab - definitely was spoiled!

@tmanbravo 2 жыл бұрын

Those Aktas are sweet! So cool CSH has a Titan Krios!

@thebumblingbiochemist 2 жыл бұрын

@aruyukimayu1199 Жыл бұрын

I envy you! I am trying to get a column selection valve V9-Cm for 3 columns and you have the one for up to 5 columns (V9-C) :)

@thebumblingbiochemist Жыл бұрын

I miss it!

@naba099 2 жыл бұрын

I am having this problem.... When I take the sample by hamilton syringe, sample tends to leak without pushing the plunger. The plunger automatically goes downwards and leaks out my sample. So I have to put the syringe always at vertical position.. which is quite problematic. Do you face this issue too? Also please make a video on how to set the methods. I am basically a molecular biologist but as a part of the project, need to confirm interaction of some proteins.

@thebumblingbiochemist 2 жыл бұрын

Hi. Could it be you're pulling to quickly maybe? Sounds like you have pressure build-up in the syringe. Sorry but the methods really vary for different things so I'm not sure I can help with that but good luck!

@ishakgun 2 жыл бұрын

Probably piston of the syringe is a little bit heavy. There are air tight syringes of Hamilton which is suitable for gas sample. You might ise these syringes

@gopalra9040 10 ай бұрын

great

@SteSews 2 жыл бұрын

Hello, I do have a probleme with my Akta system, the UV sensor doesn't work, I can't obtain a graph with UV variation. I do see the UV variation in the little screen wich is on the machine, but there is nothing on the graph so it's very hard to know where my samples are...Does anyone have an idea on how I can fix this please let me know ! thanks

@thebumblingbiochemist 2 жыл бұрын

I don't know sorry! Did you try restarting the machine? and software

@SteSews 2 жыл бұрын

@@thebumblingbiochemist thank you for your answer :) ! I did try to restart everything..but nothing change unfortunately

@thebumblingbiochemist 2 жыл бұрын

@@SteSews Really sorry to hear that. They have really great customer support though so hope they can help!

@shm6897 Жыл бұрын

The UV modules on these instruments are notoriously prone to failure over time. Usually it’s related to a breaking of the beampath or a lamp dying. They sadly are also $$$$