Browse through our How-to videos library for comprehensive step-to-step demonstrations of purification procedures, preliminary lab set ups, and product application tips filmed by our team of scientists.
Protein Purification via Affinity Chromatography - Full Tutorial
Purification with Pre-packed Column
This step-by-step tutorial demonstrates the full purification procedure of a target protein, here mCherry-Twin-Strep-tag, via gravity flow column, including the initial buffer and column preparation steps, as well as column regeneration after purification and its’ preparation for storage until further use. Gravity flow column used in the tutorial contained high-performance Strep-Tactin®XT 4Flow® resin.
Pack Your Own Column
The protein purification resin in bulk format is optimized for column affinity chromatography rather than for batch purification and is therefore intended for packing your own gravity flow columns.
This video tutorial demonstrates how to pack the 1 ml gravity flow column on the example of high-performance Strep-Tactin®XT 4Flow® resin.
Protein Purification with Large Sample Volumes
Wet Fred is an application aid for purification of (Twin-)Strep-tag® fusion proteins via gravity flow columns containing Strep-Tactin® or Strep-Tactin®XT resins. It works on the siphon principle and is used to apply large cell culture supernatant or lysate volumes into gravity flow column. Columns cannot run dry, can be left overnight, and do not need supervision. Furthermore, no sophisticated software is necessary facilitating set up and use.
The video tutorials below demonstrate the process of setting up and using the Wet Fred device, as well as the cleaning of the Wet Fred, which is to be conducted after its use and prior to the next purification round.
Resin Regeneration and Re-use
Strep-Tactin® and Strep-Tactin®XT resins for protein purification can be regenerated and re-used 3 to 5 times without loss in performance. The proper regeneration of the column and resin activation can easily be checked with HABA. The yellow HABA solution turns red (Strep-Tactin®) or orange (Strep-Tactin®XT) upon binding to the engineered biotin binding pockets of Strep-Tactin® and Strep-Tactin®XT clearly indicating that the resin is fully regenerated. Afterwards, HABA can be removed by washing with 1x Buffer W. Once the red color has disappeared the column can be re-used. If the biotin binding pocket is blocked or damaged no color shift occurs and the resin cannot be re-used.
The videos below show the respective color shifts for different resins.