Biotechniques | Basics of Making His-Tags & Nickel Affinity Chromatography
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- เผยแพร่เมื่อ 8 มี.ค. 2019
- 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.
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 ❤️😭
4th year bio-chem major here and this was very helpful in understanding the basic idea.
so insightive
Wow, this is an awesome explanation; I've been seeking for something like this for a decade!!!!, Thank you very much.
This is very useful! Thanks for uploading this video. It's exactly what I was looking for 😊💙
I've been searching for hours and finally got to watch your video. This helped me a lot in my lab! Thank you!
Glad this was helpful.
@@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?
Very well explained. Thank you so much!
Very good explained! Thank you and keep going.
Thank you so much for such a nice and simple explanation
Thanks a lot again Kevin!!!
Good good job ...thank you Dr
Amazing biology informations
Awesome explanation sir😊👌
Clear interpretation for a newer to this experiment.
wonderful lecture. Thanks.
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. :)
No problem! And thanks.
Thank you for those inforamtions ...
Thank you so much!
Thanks a lot man
Amazing 👏 🙀
Thank u so much for making this vedio with ausome explanation
You're very welcome!
It is really helpful!
Thank you!
Excellent explanation. It would be great if you added little details about how to make the protein (induction, SDS, etc...).
it will be great if you can show how to design the primers for creating these vectors.
important to say, that the his-tag does not undergo post-transcriptional modifikation , or did I miss something? VERY HELPFUL VIDEO!!
When wash with salt solution that nickel binded to NTA is removed or not sir?
How about the antibiotic resistance gene?
what would be the procedure, if I want to introduce his tag
protein into a cyanobacteria
what if a protein contained histidin in the sample and is not our protein of interest?
in the gel analysis, what are the weights on the molecular ladder?
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).
Buen video bro, lastima que entendí la mitad porque no sé ingles jajaja pero estuvo muy bueno
Vector + Gene = Plasmid
Abdala Covid vaccine brought me here