Affinity Chromatography Explained

This video introduces affinity chromatography, an essential laboratory method to isolate target biomolecules from complex mixtures. Whether you're a student, researcher, or simply curious about the science behind this technique, this video is for you!
First, we show you how it works by taking advantage of the specific binding interactions between a target biomolecule and a complementary ligand immobilized on a stationary phase. Then, we explain the different components of the stationary phase. Finally, we walk you through the various steps of an affinity chromatography experiment. As an example, we use Immobilized Metal Affinity Chromatography (IMAC), one of the most common kinds of affinity chromatography and a method we apply in our laboratory daily.
By the end of this video, you'll understand how to use affinity chromatography to isolate and purify target biomolecules from complex mixtures. However, if you would like to go into more depth on the topic, check out the video linked below:
cube-biotech.c...

[@supermunheeb]

I do this In my lab with his-tag for bacterial pesticidal proteins. this explanation helps clear things up. if im not mistaken the reason we conduct dialysis buffer exchange after is to remove any excess imidazole from our target protein as it is toxic to the insects we test and may interfere.

[@CubeBiotech]

Glad that we could help. And yes this sounds reasonable. Normal Imidazole concentrations in His-tag elution buffers is 500µM.
This almost certainly must be washed away first through dialysis for any downstream animal tests.

[@vanesa5275]

such a cute video and great explanation!! thank you

[@CubeBiotech]

Thank you :)

[@rolfandco.4766]

such a cool and interesting video! NEVER KNEW THIS!

[@CubeBiotech]

Glad that we could help you there. Did you mean something specifically?

[@rolfandco.4766]

@@CubeBiotech just the whole process in general! I had to do an oral presentation and this was one of the main topics!

[@safiraandeetasaree]

Great video! Love the animation so muchh!

[@CubeBiotech]

Thank you very much as well :)
Glad to see that the effort pays off.😊

[@agoldendream]

studying for my MCAT thank you!

[@CubeBiotech]

Good Luck !

[@lucianehernandez2778]

So helpful!

[@CubeBiotech]

Thank you :)

[@beatrizcalasensedecampos6991]

Super cute and helpfull, thank you! ❤

[@CubeBiotech]

Thank you :)

[@056vatsalapandey3]

can someone explain me the affinity and specificity logic

[@CubeBiotech]

It is a trade-off. Higher affinity means higher vield, but that usually comes with less purity/specificity. Because if you make it easier to bind, false targets also get a chance to bind.
And in the other direction this applies as well. If you make it harder to bind - higher specificity, you get purer results but also less yield.

[@favourndakara]

thank you

[@CubeBiotech]

You are welcome :)

[@mariagiselsaldanapintor4229]

I love it

[@CubeBiotech]

We are glad that you like it :)