µMACS and MultiMACS His Isolation Kits

µMACS and MultiMACS His Isolation Kits

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

Detailed procedure

After cell lysis His-tagged proteins are magnetically labeled with µMACS Anti-His 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 His-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 His Isolation Kits facilitate the isolation of His fusion proteins from different sources. The super-paramagnetic µMACS MicroBeads included in the kits are conjugated to an anti-His monoclonal antibody, enabling fast and effective magnetic labeling of His-tagged fusion proteins. The protocol allows for protein isolation within just 1.5 hours. No centrifugation or buffer removal steps are required.

Downstream applications

The (co-)immunoprecipitated proteins can be analyzed by SDS-polyacrylamide gel electrophoresis, Western blot analysis, or mass spectrometry
1
. Enzymatic on-column assays can also be performed using the thermoMACS Separator.
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