µMACS DYKDDDDK Isolation Kit130-101-591
产品说明
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 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.
特点
Fast: from cell lysis to pure protein in just 1.5 hours
Convenient: avoid multiple centrifugation and buffer removal steps
Sensitive: enables working with limited starting material
相关图片
Principle of MACS Technology for the isolation of epitope-tagged proteins