His tagged protein purification from E Coli
Автор: Jonathan's Enzymes
Загружено: 2025-08-21
Просмотров: 994
This video demonstrates how to purify a His tagged protein from E. Coli lysate using nickel agarose (Ni-NTA). I use BioRad Econocolumns for this, but with a syringe to push washes through faster (This makes purification much easier!). I also show how to use a Nanodrop spectrophotometer to estimate the protein concentration of each fraction as it elutes from nickel agarose.
The steps of the protocol are as follows:
1.) Choose the right amount of nickel agarose and equilibrate in water.
2.) Mix nickel agarose with E. Coli lysate for 15 minutes.
3.) Wash off contaminating proteins / DNA in the Econocolumn.
4.) Elute the His tagged protein using imidazole.
5.) Measure the concentration of protein in each elution fraction.
6.) Continue with dialysis, anion exchange, or SDS-PAGE followup.
I hope you enjoy this video. If you want to learn more about Solidzymes and the purification services we offer visit solidzymes.com!
https://solidzymes.com/
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