The Magnetic Phospho Enrichment Kit combines the phosphospecificity of our TALON-based Phosphoprotein Enrichment Kit with the convenience of magnetic bead separation to provide a simple, rapid, metal affinity-based method for isolating microgram quantities of phosphorylated proteins from mammalian cells and tissues. Magnetic particles in the beads facilitate quick and easy purification of microscale quantities of phosphoproteins when placed on a magnetic separator.
Isolate Cytosolic and Membrane-Bound Proteins
Each kit supplies a complete set of buffers along with our Phospho Magnetic Beads for group-specific enrichment of all types of phosphoproteins, both cytosolic and membrane-bound, that contain a phosphorylated amino acid side chain—including serine, tyrosine, or threonine.
The extraction/loading buffer contains a non-ionic detergent to extract cellular proteins. Wash and elution buffers are detergent-free. If desired, phosphoproteins can be eluted in pH 9.0 buffer, which is more suitable for downstream applications.
Microscale Purification in Small Elution Volumes
Phospho Magnetic Beads are supplied as a 5% suspension, with a demonstrated binding capacity of 400 µg of alpha-casein per ml of suspension. Different amounts of beads may be used, depending on the initial sample concentration. Phosphoproteins can be eluted in small volumes (50–200 µl) to yield concentrated samples.
Highly Selective Enrichment of Phosphoproteins
Only a small percentage of all cellular proteins are phosphorylated, so it is often necessary to enrich for this fraction before beginning an analysis. The TALON PMAC Magnetic Phospho Enrichment Kit can be used with any mammalian cell type or tissue sample. The yield of phosphorylated protein varies with different cell lines. The enrichment procedure is highly efficient.