The group of products for the isolation of epitope-tagged proteins consists of µMACS and MultiMACS™ Isolation Kits for the most common epitopes, i.e., GFP, HA, c-myc, GST, His, and DYKDDDDK (also known as FLAG®-tag). The isolation principle is based on superparamagnetic MicroBeads conjugated to the respective epitope-specific antibodies. The MicroBeads are incubated with the cell lysate and bind instantly to the target protein. The sample is loaded onto a µMACS Column placed in the magnetic field of a µMACS Separator. The magnetically labeled epitope-tagged and associated proteins are retained on the column during the washing steps and can be eluted either under native or denaturing conditions. The µMACS Epitope Tag Isolation Kits were developed for manual, low-throughput applications in combination with the µMACS Separator. The procedure can easily be up-scaled with the MultiMACS Epitope Tag Isolation Kits to a semi- or fully automated, flexible processing of 8–96 samples in parallel by utilizing the MultiMACS M96/M96thermo Separators.
Following elution the GFP-, HA-, c-myc-, His-, and DYKDDDDK-tagged fusion proteins can be detected using Miltenyi Biotec’s monoclonal antibodies that are specific for these epitopes. All antibodies are available conjugated with horseradish peroxidase for Western blot or ELISA applications. In addition HA, c-myc, His, and DYKDDDDK antibodies are coupled to fluorescent dyes (i.e. FITC, PE) for direct analysis using fast and convenient flow cytometry and fluorescent microscopy. Biotin-conjugated antibodies are also available for indirect fluorescent labeling.
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