For expressing, purifying, and detecting
proteins carrying a Strep-tag® II or a 6xHis tag
and a Strep-tag II
Two-step protein purification system
His·Strep pQE-TriSystem Vector Set
pQE-TriSystem Strep Vector
Strep-Tactin® Superflow and Superflow Cartridges
Strep-Tactin Magnetic Beads
Strep-tag Antibody

Contents
Kit Contents 4
Storage and Stability 4
Product Use Limitations 5
Quality Control 5
Technical Assistance 5
Product Warranty and Satisfaction Guarantee 6
Safety Information 6
Introduction 7
Construct screening for the best expression template 8
Purification scale 9
Buffer and reagent compatibility 9
Protocols
Growth of E. coli Cultures (50 ml) 10
Preparation of Cleared Lysates from E. coli Cell Cultures 11
Preparation of Cleared Lysates from Baculovirus-Infected Insect Cells 13
Preparation of Lysates from Transfected Mammalian Cells 15
Manual Purification of Strep-tagged Proteins Using a Syringe 17
Purification of Strep-tagged Proteins Using Liquid Chromatography Systems 18
Batch Purification of Strep-tagged Proteins Using Strep-Tactin Superflow 20
Micro-Scale Purification of Strep-tagged Proteins Using Strep-Tactin Magnetic Beads 21
Immunodetection using the Strep-tag Antibody (Chemiluminescent Method) 23
Immunodetection using the Strep-tag Antibody (Chromogenic Method) 25
Troubleshooting Guide 27
Appendix A: Two-Step Purification Using a His-tag and a Strep-tag 32
Protein Purification from Insect-Cell Growth Medium 35
Manual Purification of His-tagged Proteins Using a Syringe 36
Purification of His-tagged Proteins Using Liquid Chromatography Systems 37
Batch Purification of His·Strep-tagged Proteins Using Ni-NTA Matrices 38
Micro-Scale Purification of His·Strep-tagged Proteins Using Ni-NTA Magnetic Agarose Beads 39
Appendix B: Composition of Buffers 41
Appendix C: Regeneration of Strep-Tactin Superflow 45
Ordering Information 46