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LC-MRM Quantification of Src-Family Kinases using Protein Standards Expressed in Cell-Free Systems

Promega Corp.

LC-MRM Quantification of Src-Family Kinases using Protein Standards Expressed in Cell-Free Systems

6 Jul 2015

This poster describes a method for the quantification of SFK expression using peptides unique to each sequence. Each of the SFK proteins screened in this study produced at least 2 unique peptides, and all SFK proteins could be detected and quantified in a single experience when applied in parallel, offering a clear advantage over immunoblotting.

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HaloTag® Mammalian Protein Purification System

Promega Corp.

It can be challenging to purify sufficient high-quality protein from mammalian cells. Boost protein recovery 30-60% with an optimized covalent protein tag, called HaloTag®. This complete kit offers strong improvements in recovery, purity, cost and protein functionality over historical FLAG® and poly-His tag purification strategies.

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Promega Corp.

  HaloLink - Promega Magnetic Beads for Covalent immobilization of HaloTag® fusion proteins.   Features of the Promega HaloLink Magnetic Beads: • Magnetic-Based: Easy and quick immobilization of HaloTag® fusion proteins with minimal loss of sample material. • Covalent Bond: Allows stringent washing of immobilized HaloTag® fusion protein, removing nonspecific proteins and enhancing overall data.  • Multiple Applications: Immobilized HaloTag® fusion proteins can be used for protein interaction analysis, enzyme assays and purification of fusion proteins containing a protease cleavage site. HaloLink™ Magnetic Beads from Promega provide a rapid and reliable method to covalently capture and immobilize HaloTag® fusion proteins to a paramagnetic particle. Immobilization through the HaloTag® Ligand provides for consistent, surface-directed orientation of the fusion protein, thus providing more reliable results in applications such as protein:protein interaction studies with the fusion protein. The result is a paramagnetic bead with very high binding capacity for HaloTag® fusion proteins and very low nonspecific protein binding. The paramagnetic feature allows for streamlined assay development and facilitates applications with automated assays developed on robotic platforms. A variety of vectors for the expression of HaloTag® fusion proteins in bacterial, mammalian or cell-free systems are available.

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