Video Tutorials
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.
Protein purification via magnetic beads
In addition to 50% suspension, Strep-tag® products for protein purification come in variety of other formats allowing various applications.
Magnetic beads are a versatile tool suitable for many applications, such as batch purification of many samples in parallel, pull-downs, immunoprecipitation or co-immunoprecipitation.
This step-by-step tutorial demonstrates the full purification procedure of a target protein, here mCherry-Twin-Strep-tag, via magnetic beads (MagStrep Strep-Tactin®XT beads).
Protein purification with spin columns
For small-scale experiments with a limited number of samples, spin columns are a convinient tool. They are perfect for manually purifying a number of different proteins at the same time in a fast and efficient way.
This step-by-step video tutorial demonstartes the full purification procedure of target proteins using Strep-Tactin®XT 4Flow® high capacity Spin Column Kit, reaction tubes, vortex and centrifuge.
Resin Regeneration and Re-use
Strep-Tactin® and Strep-Tactin®XT resins for protein purification can be regenerated and re-used >10 times* without loss in performance. 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 100 mM NaOH. Once the red color has disappeared and additional washing with Buffer W was performed to remove NaOH, 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.
*under ideal conditions >50 regeneration cycles are possible
The videos below show the respective color shifts for different resins.
Cell isolation
Cells expressing Strep-tagged surface proteins can be selected via these affinity tags through various methods such as flow cytometry, magnetic separation, or affinity chromatography. Alternatively, Strep-tagged proteins that bind to cells can be combined with different Strep-Tactin® and Strep-Tactin®XT conjugates. This creates selection reagents that can be used to isolate specific cell populations.
The following video tutorials show all the steps of cell isolation from various samples using affinity chromatography with Strep-Tactin® TACS Agarose Columns and magnetic cell isolation using Strep-Tactin® Magnetic Microbeads, both in combination with a Strep-tagged Fab fragment.
