Phosphopeptide Enrichment Spin Columns
Phosphopeptide Enrichment Spin Columns
Brand: Takara Bio.
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Phosphopeptide Enrichment Spin Columns
Phosphopeptide Enrichment Spin Columns
Our Phosphopeptide Enrichment Spin Columns and Phosphopeptide Enrichment Buffer Kit can enhance detection of phosphorylated peptides that would otherwise be undetectable. The straightforward protocol enriches your protein digests prior to detection by mass spectroscopy or HPLC.
The spin columns contain a unique immobilized metal affinity chromatography resin that binds phosphopeptides. These columns have the capacity to bind up to 250 µg of phosphopeptide and can accommodate up to an 850 µl sample volume. The columns fit into most microcentrifuges. There is no need to pre-equilibrate—simply spin out the storage buffer.
Our buffer kit provides optimized loading and elution buffers for use with the spin columns and is recommended for optimal results. Each buffer kit includes enough buffer for use with 25 spin columns and saves time when compared to making your own buffer.
Our Phosphoprotein Enrichment Kit provides an effective affinity-based procedure for isolating phosphorylated proteins from mammalian cells and tissues.
Overview
- Efficient, specific enrichment for all types of phosphopeptides
- Convenient prepackaged columns
- Enrich for any type of phosphopeptide—phosphotyrosine, phosphoserine, or phosphothreonine
- Obtain increased sensitivity in mass spectroscopy analysis
Applications
- Phosphopeptide enrichment
Phosphopeptides of beta-casein purified using the Phosphopeptide Enrichment Spin Columns
Phosphopeptides of beta-casein purified using the Phosphopeptide Enrichment Spin Columns. Beta-casein protein was digested with trypsin (sequencing grade). The protein digest was diluted at a ratio of 1:1 with Loading Buffer and run on a Phosphopeptide Enrichment Spin Column. The enriched fraction was eluted with Elution Buffer, then freeze-dried and reconstituted in 0.1% TFA in water (v/v). Reverse phase HPLC (RP-HPLC) data is shown for the crude protein digest (Panel A), eluate (purified phosphopeptides) (Panel B) and eluate of a dephosphorylated sample (Panel C). The eluted fractions were separated by RP-HPLC on an XTerra RP18 column (5 μm, 4.6 x 150 mm) with UV detection at 215 nm (Solvent A: 0.1% TFA in water [v/v], Solvent B: 0.1% TFA in acetonitrile [v/v]). The two phosphopeptide fractions collected in Panel B were analyzed by MALDI.
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Phosphopeptides of beta-casein purified using the Phosphopeptide Enrichment Spin Columns
Phosphopeptides of beta-casein purified using the Phosphopeptide Enrichment Spin Columns. Beta-casein protein was digested with trypsin (sequencing grade). The protein digest was diluted at a ratio of 1:1 with Loading Buffer and run on a Phosphopeptide Enrichment Spin Column. The enriched fraction was eluted with Elution Buffer, then freeze-dried and reconstituted in 0.1% TFA in water (v/v). Reverse phase HPLC (RP-HPLC) data is shown for the crude protein digest (Panel A), eluate (purified phosphopeptides) (Panel B) and eluate of a dephosphorylated sample (Panel C). The eluted fractions were separated by RP-HPLC on an XTerra RP18 column (5 μm, 4.6 x 150 mm) with UV detection at 215 nm (Solvent A: 0.1% TFA in water [v/v], Solvent B: 0.1% TFA in acetonitrile [v/v]). The two phosphopeptide fractions collected in Panel B were analyzed by MALDI.
