Meet the AKTA! (and don't be intimidated by it) - Fast Protein Liquid Chromatography (FPLC)
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- čas přidán 29. 06. 2021
- 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
For a longer version with more tips: • Meet the AKTA! Fast Pr... - Věda a technologie
Good video 🙂 ! I would really appreciate if you can make one quick video when you actually are purifying the sample. It will also make us clear how to actually operate it and control the software as well ! I am sure there are many who will want to see that :) .🙏
Thanks! Maybe in the future
@@thebumblingbiochemist Really agree with him, if you can make a quick video about protein purification from the start to end and it will be great. Thanks😄
Hi - if you go to the protein purification playlist you will find more practical videos: czcams.com/play/PLUWsCDtjESrEtqQkEXsQpTyRfMhywBVQ7.html including this one I made based on your feedback so thanks for the suggestion! czcams.com/video/rwpgoL86t_Q/video.html
This was so helpful!! Thank you so much.
Glad I could help!
AKTA is Swedish for “Pure” - comes from the days of Pharmacia. Hence - Sephadex - Separation Pharmacia Dextran & Sepharose - Separation Pharmacia Agarose. Pharmacia was the inventors of the first LC columns and resins.
Cool! Thanks for sharing!
Can you explain what the UV detector does? If we are only keeping track of the time it takes for molecules to exit the column,then why do we need a UV detector?
The UV detector shows us when protein is coming off the column. Then we can collect the fractions corresponding to that time.
@@thebumblingbiochemist thanks for your response,could you also explain why a PH valve is necessary?
The pH reader is optional. It can be a good check if you have an issue to see if the pH of the buffer was okay. And if you're doing a pH gradient, you can monitor that. But it isn't required
@@thebumblingbiochemist thank you again.
liked and subscribed.
What is the effect after joining two hisTrap columns rather than one?
You increase your binding capacity - but remember to double your buffer volumes
@@thebumblingbiochemist thx, but will the speed increase and the purity increase too?
no
Unfortunate that the video is mirror-imaged. :(
Sorry - please check out this later video: czcams.com/video/KywmjA3uucA/video.html