The µMACS and MultiMACS DYKDDDDK Isolation Kits facilitate the isolation of DYKDDDDK-tagged (also known as FLAG
®
-tagged) proteins from different sources. The super-paramagnetic µMACS MicroBeads included in the kits are conjugated to an anti-DYKDDDDK monoclonal antibody, enabling fast and effective magnetic labeling of DYKDDDDK-tagged fusion proteins. The protocol allows for protein isolation within just 1.5 hours. No centrifugation or buffer removal steps are required.

Data and images for µMACS™ and MultiMACS™ DYKDDDDK Isolation Kits

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Principle of MACS Technology for the isolation of epitope-tagged proteins.

Figure 1

Principle of MACS Technology for the isolation of epitope-tagged proteins.

Specifications for µMACS™ and MultiMACS™ DYKDDDDK Isolation Kits

Overview

The µMACS and MultiMACS DYKDDDDK Isolation Kits facilitate the isolation of DYKDDDDK-tagged (also known as FLAG
®
-tagged) proteins from different sources. The super-paramagnetic µMACS MicroBeads included in the kits are conjugated to an anti-DYKDDDDK monoclonal antibody, enabling fast and effective magnetic labeling of DYKDDDDK-tagged fusion proteins. The protocol allows for protein isolation within just 1.5 hours. No centrifugation or buffer removal steps are required.

Detailed product information

Detailed procedure

After cell lysis DYKDDDDK-tagged proteins are magnetically labeled with µMACS Anti-DYKDDDDK MicroBeads. The sample is loaded onto a MACS Column placed in the magnetic field of a µMACS or thermoMACS Separator in which the magnetically labeled DYKDDDDK-tagged and associated proteins are retained during the washing steps. After elution with denaturing elution buffer, the precipitated proteins can be analyzed by SDS-PAGE and Western blot. Alternatively, native proteins can be eluted by pH-shift using triethylamine pH 11.8 or by removing the column from the magnetic field and applying a suitable native elution buffer.

Applications

The µMACS and MultiMACS DYKDDDDK Isolation Kits facilitate direct immunoprecipitation of DYKDDDDK fusion proteins from cell lysates at an analytical scale. They can also be used in co-immunoprecipitation experiments for the isolation of associated proteins or protein complexes in protein-protein interaction studies.

Downstream applications

The (co-)immunoprecipitated proteins can be analyzed by SDS-polyacrylamide gel electrophoresis, Western blot analysis, or mass spectrometry. Enzymatic on-column assays can also be performed using the thermoMACS Separator.

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µMACS™ and MultiMACS™ DYKDDDDK Isolation Kits

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