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Unicorn Stuff
United States
Registrace 30. 01. 2023
I make chromatography tutorials using Unicorn software, mostly.
I have a Ph.D. in Biochemistry and I've been working with Unicorn for 20 years, since Unicorn 3.11.
I have a Ph.D. in Biochemistry and I've been working with Unicorn for 20 years, since Unicorn 3.11.
How Fast Forward Through Methods With The Next Breakpoint Instruction in Unicorn with AKTA systems.
Find out how to fast forward through Unicorn Methods on any AKTA system. In this method we simulate a power failure halfway through a chromatography run then discuss how to get back up and running using the Next Breakpoint instruction.
If you need help finding Breakpoints in Unicorn be sure to watch my deep dive into understand them: czcams.com/video/tNcAEIAwtm8/video.html
You may also like watching my video about Blocks which also discusses breakpoints: czcams.com/video/szYyu6MjOrw/video.html
For further reading generally on how to use the AKTA Avant see: cdn.cytivalifesciences.com/api/public/content/digi-10624-original
Reach me at my Linkedin page www.linkedin.com/in/chad-blamey-ph-d-649a336/
Music:
Midnight by Massobeats - freetouse.com/music/massobeats/midnightChamomile by Massobeats - freetouse.com/music/massobeats/chamomile
Lush by Massobeats - freetouse.com/music/massobeats/lush
This music is not mine it is used under a creative commons license. Thanks for letting me use your music I really like it!
If you need help finding Breakpoints in Unicorn be sure to watch my deep dive into understand them: czcams.com/video/tNcAEIAwtm8/video.html
You may also like watching my video about Blocks which also discusses breakpoints: czcams.com/video/szYyu6MjOrw/video.html
For further reading generally on how to use the AKTA Avant see: cdn.cytivalifesciences.com/api/public/content/digi-10624-original
Reach me at my Linkedin page www.linkedin.com/in/chad-blamey-ph-d-649a336/
Music:
Midnight by Massobeats - freetouse.com/music/massobeats/midnightChamomile by Massobeats - freetouse.com/music/massobeats/chamomile
Lush by Massobeats - freetouse.com/music/massobeats/lush
This music is not mine it is used under a creative commons license. Thanks for letting me use your music I really like it!
zhlédnutí: 12
Video
Timer Instruction in Unicorn for AKTA Chromatography Systems
zhlédnutí 27Před 14 dny
The Timer is one of the most helpful instructions there is. You can only use it in the System Control module while you are doing a run, either manual or a method. The instruction is available for all AKTA systems (Avant, Pure, Go, Pilot, Process and more) controlled by Unicorn. You can use it for convenience to pause the system at a specific time or you can use it as a diagnostic to check to se...
Peak Fractionation Instruction in Unicorn for AKTA system
zhlédnutí 53Před 28 dny
Here we discuss the Peal Fractionation instruction in Unicorn for use on AKTA chromatography systems like the Avant, Go or Pure. The Peak Fractionation instruction is used in the manual and method modules for some tight fraction action! This instruction will allow you to save tons of tubes as you crank out run after run. For further reading generally on how to use the AKTA Avant see: cdn.cytiva...
Gradient Instruction In Unicorn for AKTA chromatography systems
zhlédnutí 58Před měsícem
Here we discuss the Gradient Instruction in Unicorn for use on AKTA chromatography systems like the Avant, Go or Pure. The Gradient instruction is used in the manual and method modules to control the ratio of pump A and pump B. When you are working with the gradient instruction in the method editor be sure to pay careful attention to the breakpoints as these can change in two places at once. Fo...
The Secrets Of The Flow Instruction
zhlédnutí 58Před 2 měsíci
This is a video about instructions in Unicorn. My goal is to eventually describe how to use (almost) every instruction in Unicorn for use on the AKTA Avant, Go and Pilot. We'll see how far I get. Covered here is the System Flow instruction. For further reading generally on how to use the AKTA Avant see: cdn.cytivalifesciences.com/api/public/content/digi-10624-original Reach me at my Linkedin pa...
Packing A HIC Tricorn Column
zhlédnutí 169Před 3 měsíci
Here we pack a column that is 5 mm wide, so tiny, to a height of 8 cm for chromatography purification. We use a Tricorn column from Cytiva with Hydrophobic Interaction Chromatography (HIC) resin (or media). We use Phenyl HP to pack this column. You can use this technique to pack just about any Tricorn column. You could use this technique to pack any flavor of column like IEX, affinity, desaltin...
Packing The XK Column For Chromatography
zhlédnutí 534Před 3 měsíci
Hey so here we pack an 8 cm XK column from Cytiva with Hydrophobic Interaction Chromatography (HIC) resin (or media). We use Phenyl HP to pack this column. You can use this technique to pack just about any XK column, large or small. You could use it to pack an IEX, affinity, desalting or SEC column as well. I do not represent Cytiva & this is not a paid promotion. For some moderately unhelpful ...
Woohoo! Changing The Sounds Your AKTA Avant Makes With Unicorn Software.
zhlédnutí 92Před 4 měsíci
Here we totally nerd out and have some 'fun' with Unicorn by changing the sounds your system makes. This works with any AKTA chromatography system with Unicorn control, the Avant, Pure, Go, Pilot and Process just for starters. I made my system say Woohoo like Homer Simpson. Just trying to have some fun and lighten the mood maybe. For further reading generally on how to use the AKTA Avant see: c...
How To Make A Preset SEC Method On The AKTA Go
zhlédnutí 228Před 4 měsíci
In this introductory video we go over EVERY step in how to make a Size Exclusion Chromatography (SEC) method for the AKTA Go using Unicorn software from Cytiva. We also set up the system and briefly review the results. I don't represent Cytiva and I am not (nor have I ever been) paid by Cytiva. I'm just super into chromatography. How to make a custom column in Unicorn. czcams.com/video/qYlFHvSk...
A Basic Intro To Understanding Chromatography Results Collected With The AKTA Chromatography System
zhlédnutí 235Před 5 měsíci
In this introductory video I show you how to view and understand your chromatography results. It's super basic. I used Unicorn software that runs the AKTA chromatography system suits like the Avant, Pure, Pilot and Go. Here I use an affinity purification I did with Protein A resin/media. Please leave a comment below and good luck with your purifications! Reach me at my Linkedin page www.linkedi...
The Base Matrix Of Your Chromatography Resin Is Really Important.
zhlédnutí 74Před 6 měsíci
Here we talk about how the base matrix of chromatography resin (media/gel) affects performance of your runs. I mostly make videos about using the AKTA Chromatography systems like the Avant, Pure, Pilot and Go with Unicorn Software from Cytiva for preparative purification. (I don't work for Cytiva and I'm not paid by them either.) This talk also applies to some other kinds of chromatography like...
Replacing The Inline Filter Of An AKTA Avant Chromatography System.
zhlédnutí 312Před 6 měsíci
Wait the AKTA Avant has an inline filter? The inline filter often gets dirty or contaminated which can lead to high back pressure of contamination of your purified samples. Here I will show you how to replace the inline filter, it is easy and takes about 2 minutes. But honestly you don't even need the filter. I took my filter out and never looked back. I always filter my cell harvests and usual...
MultiDimensional Chromatography Setup
zhlédnutí 79Před 8 měsíci
In this video we learn how setup for the multidimensional chromatography method we made a little while back using a desalting column and some anion exchange columns on an AKTA Avant. This method allows us to test many different conditions using multiple buffers at the same time! For example the sample can be in high salt with low pH conditions and the method will still work. This is an 'advance...
An Example of Multi Dimensional Chromatography: Desalt and Ion Exchange
zhlédnutí 109Před 9 měsíci
In this video we learn how to do multi dimensional chromatography using a desalting column and some anion exchange columns on an AKTA Avant. To start we are building on an a resin selection project we did earlier see below. This method will allow us to test many different conditions using multiple buffers at the same time! For example the sample can be in high salt with low pH conditions and th...
Creating Reminders In Unicorn Chromatography Software.
zhlédnutí 42Před 9 měsíci
In this quick video I'll show you how to set up a reminder for an AKTA Avant, or Pure or any other system that is controlled by Unicorn Software. This can be convenient for reminding you when to do periodic maintenance with your system, or just to tell you to take a minute for yourself to eat lunch. I hope you enjoy. I thought this content would be helpful because I haven't found anything about...
Changing The Settings On The AKTA Avant in Unicorn. Most Helpful Thing Ever!
zhlédnutí 175Před 10 měsíci
Changing The Settings On The AKTA Avant in Unicorn. Most Helpful Thing Ever!
How to Fix a Broken Fraction Collector Tube on the AKTA Avant Chromatography System
zhlédnutí 239Před 10 měsíci
How to Fix a Broken Fraction Collector Tube on the AKTA Avant Chromatography System
How Unlock Unicorn Method or Result Files For All Chromatography Systems.
zhlédnutí 323Před 11 měsíci
How Unlock Unicorn Method or Result Files For All Chromatography Systems.
Setup For Selection of Ion Exchange Medias Using A Unicorn Method on the AKTA Avant.
zhlédnutí 158Před 11 měsíci
Setup For Selection of Ion Exchange Medias Using A Unicorn Method on the AKTA Avant.
Look At All Those Zeros! Reading Unicorn Language with AKTA Chromatography Systems
zhlédnutí 256Před rokem
Look At All Those Zeros! Reading Unicorn Language with AKTA Chromatography Systems
Best and Worst About the AKTA Avant (in my opinion)
zhlédnutí 101Před rokem
Best and Worst About the AKTA Avant (in my opinion)
Selection of Ion Exchange Medias Using A Unicorn Method on the AKTA Avant.
zhlédnutí 256Před rokem
Selection of Ion Exchange Medias Using A Unicorn Method on the AKTA Avant.
Setting Up a Custom Affinity Method Using Scouting for the AKTA Avant Chromatography System.
zhlédnutí 288Před rokem
Setting Up a Custom Affinity Method Using Scouting for the AKTA Avant Chromatography System.
Testing a Unicorn Method on the AKTA Avant Chromatography System
zhlédnutí 180Před rokem
Testing a Unicorn Method on the AKTA Avant Chromatography System
Writing a Custom Affinity Method Using Scouting for the AKTA Avant Chromatography System.
zhlédnutí 404Před rokem
Writing a Custom Affinity Method Using Scouting for the AKTA Avant Chromatography System.
Making a Custom Column in Unicorn Chromatography Software for the AKTA Avant
zhlédnutí 127Před rokem
Making a Custom Column in Unicorn Chromatography Software for the AKTA Avant
Break Points and Timing In Unicorn on AKTA Avant
zhlédnutí 314Před rokem
Break Points and Timing In Unicorn on AKTA Avant
Understanding blocks in Unicorn software for AKTA Avant
zhlédnutí 942Před rokem
Understanding blocks in Unicorn software for AKTA Avant
Adding A Variable In Unicorn Chromatography Software for AKTA Avant.
zhlédnutí 212Před rokem
Adding A Variable In Unicorn Chromatography Software for AKTA Avant.
If equilibration is same as elution, running "block" elution". I need to check if phase will work as a block. I fear double tapping next breakpoint. I have rerun a load block when the air sensor was triggered.
I'm not sure of your exact situation, but the first step is to carefully reread the method. You can either restart the method and you next breakpoint to quickly get you back where you need to be. The other option with out restarting the method is to use the "block" instruction to just re-read in the elution block where the air sensor prematurely triggered. Gook luck! btw I haven't covered the "block" instruction yet.
I am using AKTA Avant and AKTA Pure 150.
Nice! I have those systems too.
Hello thanksss
I'm glad you liked the video!
is it really neccessary to do the pump wash for every run in ÄKTA pure / ÄKTA avant, 'cause I see that they always do a short pump wash when switch to other inlet possition?
Yes and no. I would say you want to do a pump wash every time you start a new run, but not every time you switch to a new inlet position. So say you start a run by washing inlets 1 and 2 you don't have to rewash them later in the middle of the run to switch from one inlet to the next. UNLESS the solution in one of the inlets would totally muck up the run some how, like in affinity chromatography, for example you don't want to have Ni anywhere near the column when loading you sample with a His-tag (HisTrap) column.
thank you for your terrific videos!
nice video, thanks!
Glad you liked it!
👍
THANK YOU MY FRIEND .
I'm glad you liked this video! I worked really hard on it and to find that someone found it useful is great!
Thank you so much. do you have two step purification, protein A and desalt column training one? usually I need buffer exchange after purification to remove the tris component which was used neutralized the elution buffer. thanks.
Do you know how much about Akta Training? I am looking for a protein related jobs now...
thank you for you terrific work!
Thanks for watching!
thank you for you method!
I have been using a pause at the end of the run. The chromatogram does not like this. Peak shifts to the left changing from volume to time.
I have a whole series of videos about navigating modules so you can get the chromatogram you are looking for. They were some of the first videos I did, if you haven't seen them I'd recommend checking them out, czcams.com/video/m5kp9wM63ZQ/video.html
I am getting 0.mg/mL concetration even after providing the UV path length and ext.co effecient value. Please help me am i missing anything
It's hard to know what's going on with the information you provided. Tell me more or message me a snap shot of your chromatogram. Here are some guesses about what I think is going on. Did you integrate the peak? The area integrated needs to just be the peak of interest. Be sure the baseline is properly drawn/calculated, you can really only adjust the baseline in the classic evaluation module (so far as I know). Sorry I didn't answer quickly...
Thanks for the very helpful video. Do you equilibrate the slurry with water before or after you add them to the column?
Good question. Ideally you should do it before, the slurry will settle very quickly if it is fully equilibrated, but if you don't mind waiting for it to settle then you can do it while it is on the column.
Some possible ideas for future videos....1) Pressure measurement in ÄKTA Systems 2) Setting delay volumes in ÄKTA Systems. 3) Column care and maintenance (Superdex Increase) Thanks!
Thanks for the idea!
Nice video! How do I see all settings in an already saved method?
Click on the text tab in the middle of the screen, go to 2:43 mark in this video to see what I'm talking about. You'll have to read through the method to find the setting you want. Once you find the setting be sure to press the change button, then save. Good luck.
Nice video! Adds some fun to lab work. I can never remember how to change pressure units between mPa and psi in unicorn 7. Can't find it in the user manual. Any thoughts?
Thanks, I'm glad you liked the video! You can change the unit for results. In any of the modules go to Tools in the menu bar and select Options. Then in the window that pops up select the View tab, towards the bottom of the tab there is an option for Pressure Unit. It doesn't look like you can change the units in the Method Editor or System Control, only for results. It's a good question though.
Thanks for sharing your knowledge in a really clear and helpful way!
Glad it was helpful! I don't know why this video was such a challenge to make, probably because everyone is going to have something different, so I'm glad you liked it enough to comment.
Hi, can you upload column-packing videos??
I hope to soon. I have so many ideas and each video takes a bit to make. Is there a kind of column in particular you want to see packed?
HIC
Done! I did one, I should have some more up sometime in the near future. Not making any promises. czcams.com/video/z02_8-yjmOk/video.html
These videos are super helpful. I don't think there are any other resources out there like this for AKTA. I was wondering, if you have a loop valve with 5 loops, can you run scouting using five different inlets that each would store different proteins in each of the five loops. Then each loop could be desalted one by one? The first step would be IEX and the second step would be desalting.
Yeah you could, you'd either need two valves plumbed like the outlet valve or one like the column valve. There's no reason you couldn't have a second column valve you'd just have you use your imagination when it came to understanding the flow path.
Thank you for such a nice explaination, i am new to Äkta system and this helps me a lot :)
Glad it was helpful! Really means a lot! I'm trying to gear this channel towards people, such as yourself, that are new to the Avant and other ÄKTA systems.
☺@@UnicornStuff Wish your channel gets more views and more people learn from here. Surprisingly, when I search for Äkta, the recommendation did not come directly; it only came up when I searched with Unicorn. Such a nice channel.❤
Thank you! This was very helpful, as we had some ancient runs which we could not delete…
What really! I never imagined someone would actually use my content. So excited my videos actually fill a need. Cheers!
Thank you for sharing, it's really helpful.
Glad it was helpful! It can be really helpful to get the hang of Unicorn and the Avant at first.
Hi there, Why do you have two peaks when doing step elution?
Good question. There could be a variety of reasons all of which could be true in this case. There could be impurities in the sample that elute later. There could one target protein, but it is heterogeneous because of post transnational modifications. I believe the best answer however is the concentration of salt wasn't strong enough to get all the protein off for that step. The solution would be to increase the step length or concentration a little bit. Sorry for the late reply.
Thank you so much! It really helps me a lot.
You're welcome! I'm planning on making more videos.
Excellent teaching ! Learning something new every time 👍
Glad to hear that!
why not use sodium hydroxide for CIP?Thanks
Great question. Most protein A type resins will foul if you use >10 mM NaOH because the ligand is unstable in higher concentrations. There are some NaOH resistant resins MabSelect (Cytiva) and Toyopearl (Tosoh) are my two favorite.
question: when purgae pump head, is it necessary to turn on system flow? Thanks
No it is not, one of the check valves is always open. It is necessary to pump wash the system afterwards regardless. Thanks for your question.
Excellent teaching!thank you so much !
Glad you liked it!
Excellent teaching!👍thank you!
Glad it was helpful!
Thank you so much for the teaching 👍🙏it is very helpful ! by the way ,the back ground music is a little bit noisy if you don’t mind me saying !
Thanks for your feedback. I'm working on using the audio editor more. I'd like to have background music to cover up some of the breathing noises I make. Hopefully we find something we both like.
@@UnicornStuff I see. Understand! Thank you so much! I
Very useful , very clear explanation for a beginner ! thank you so much !any more in this Unicorn software?
Thanks! Yes I'm making more & super excited you found my channel!
@@UnicornStuff looking forward to seeing more about how to use the unicorn software , methods editor and results evaluation 🙏👍