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

[@sunburntsatan6475]

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]

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

[@nicholaskampa2993]

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

[@thebumblingbiochemist]

So happy I could help! Have fun purifying!

[@sakitlaatigue]

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]

Happy I could help! Good luck with your purifications!

[@JackieLiu0406]

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

[@thebumblingbiochemist]

HHMI structural biology lab - definitely was spoiled!

[@aruyukimayu1199]

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]

I miss it!

[@tmanbravo]

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

[@thebumblingbiochemist]

:)

[@שגיתזיוספר]

Great explanation! Loved it, thank you!

[@thebumblingbiochemist]

Glad it was helpful!

[@056vatsalapandey3]

What college did you record this

[@thebumblingbiochemist]

Cold Spring Harbor Laboratory

[@halilojaallways7204]

You Rock!! Very useful Information

[@thebumblingbiochemist]

Thank you! So glad it was helpful!

[@naba099]

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]

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]

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]

great

[@SteSews]

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]

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

[@SteSews]

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

[@thebumblingbiochemist]

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

[@shm6897]

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 $$$$