Biotinylation Case Studies

Listed below are several protein biotinylation case studies. For more details regarding our custom protein production services, click here. If you would like to browse the workflow utilized by our team in our biotinylation process, click here.

Site-specific biotinylation of Custom Protein

  • Strategy: 

    • Construct and express target protein with AviTag at the N terminus

    • The avi-tagged target protein is biotinylated in vitro by incubation with BirA ligase and biotin

    • Remove free biotin by G-25 column

  • QC: 

    • Purity >95% by SEC-HPLC

    • Pierce HABA biotin quantitation: ~1 biotin molecule per biotinylated protein

Chemical biotinylation of Custom Protein

  • Strategy: 

    • Construct and express target protein with His tag at the C terminus

    • Incubate target protein with EZ-Link NHS-PEG12-Biotin

    • Remove free biotin by G-25 column

  • QC: 

    • Purity >90% by SEC-HPLC

    • Pierce HABA biotin quantitation: ~4.5 biotin molecule per biotinylated protein