Biotechniques | Basics of Making His-Tags & Nickel Affinity Chromatography

No video

Biotechniques | Basics of Making His-Tags & Nickel Affinity Chromatography

Рет қаралды 39,319

Күн бұрын

As always, the steps of putting a gene insert into vector/plasmid AND nickel affinity chromatography depend on your specific lab and protocol. Here, we look at the general theory of engineering a poly-His tagged protein and Ni affinity chromatography and what they are used for in the lab.

Пікірлер: 38

@giftymammen1515 3 жыл бұрын

Wow, whatta explanation. I've been searching for the literature for hours and have been pounding my brain ever since on this topic. The minute I landed onto this video, all my confusions are gone, crystal clear now. Thankyou so much ❤️😭

@satinderjit4 3 жыл бұрын

4th year bio-chem major here and this was very helpful in understanding the basic idea.

@robertadu-agyei7616 3 жыл бұрын

so insightive

@AL-eh3jx 4 жыл бұрын

I've been searching for hours and finally got to watch your video. This helped me a lot in my lab! Thank you!

@CatalystUniversity 4 жыл бұрын

Glad this was helpful.

@AL-eh3jx 4 жыл бұрын

@@CatalystUniversity I have a question and would appreciate your help! If we use Nickle binding buffer containing imidazole( low concentration) before the incubation with resin, would any binding between the His-tagged protein and Ni ions happen at this stage?

@qamarhennawi9137 2 жыл бұрын

Wow, this is an awesome explanation; I've been seeking for something like this for a decade!!!!, Thank you very much.

@yourfuturedocburenbeiya 4 жыл бұрын

This is so helpful ; thank you for posting this video doc! We're using it in lab to isolate dmAANATA and I couldn't find the molecular weight but came across this video. :)

@CatalystUniversity 4 жыл бұрын

No problem! And thanks.

@jimmyyoung1657 3 жыл бұрын

Clear interpretation for a newer to this experiment.

@pingr.8730 3 жыл бұрын

This is very useful! Thanks for uploading this video. It's exactly what I was looking for 😊💙

@robertstadler9897 2 жыл бұрын

Very good explained! Thank you and keep going.

@saherk5021 3 жыл бұрын

Thank you so much for such a nice and simple explanation

@snobbips5107 5 жыл бұрын

Awesome explanation sir😊👌

@xuehanxu8814 3 жыл бұрын

Very well explained. Thank you so much!

@bioscienceswithshahtareenswati 2 жыл бұрын

Amazing 👏 🙀

@titoflash1212 3 жыл бұрын

Thanks a lot again Kevin!!!

@khurshedakabirov8671 3 жыл бұрын

Excellent explanation. It would be great if you added little details about how to make the protein (induction, SDS, etc...).

@khaledkourbane9182 4 жыл бұрын

Good good job ...thank you Dr Amazing biology informations

@davalcom 2 жыл бұрын

Thanks a lot man

@NehaSharma-kp8op 4 жыл бұрын

Thank u so much for making this vedio with ausome explanation

@CatalystUniversity 4 жыл бұрын

You're very welcome!

@uzoechisamuel 2 жыл бұрын

wonderful lecture. Thanks.

@izzatiauni3721 4 жыл бұрын

It is really helpful!

@CatalystUniversity 4 жыл бұрын

Thank you!

@carlostettey2889 2 жыл бұрын

it will be great if you can show how to design the primers for creating these vectors.

@khaledkourbane9182 4 жыл бұрын

Thank you for those inforamtions ...

@AA-gl1dr 3 жыл бұрын

Thank you so much!

@karl-leopoldkontrus6544 4 жыл бұрын

important to say, that the his-tag does not undergo post-transcriptional modifikation , or did I miss something? VERY HELPFUL VIDEO!!

@ying2694 3 ай бұрын

How about the antibiotic resistance gene?

@christinamrad7341 Жыл бұрын

what if a protein contained histidin in the sample and is not our protein of interest?

@jenna9992 4 жыл бұрын

in the gel analysis, what are the weights on the molecular ladder?

@CatalystUniversity 4 жыл бұрын

To be perfectly honest, it depends on your ladder. If you know the company and the exact product, the company website will have this available. For example, if I knew I was using the 1 kb DNA Ladder made by New England Biolabs (very good company, by the way), I could type into google search "New england biolabs 1 kb ladder". Long story short, you would need to find the exact product or at least know the size of the ladder (e.g., 1 kb).