Affinity enrichment of ATP-binding proteins The ProteoEnrich™ ATP-Binders™ Kit allows group separation of protein kinases and other ATP binding proteins, yielding partially purified cell extracts enriched in protein kinases that retain enzymatic activity. Based on an affinity resin containing immobilized ATP, the method is compatible with 2D electrophoresis, SDS-PAGE/tandem mass spectrometry, Western blot analysis, or activity assays. The unique affinity resin contains ATP covalently linked through the g-phosphate, presenting an ideal configuration for recognition by the conserved ATP binding pocket of kinases. The kit contains the ATP-polyacrylamide-based resin, spin filters, and optimized buffers required to process extracts prepared from crude mammalian cell or tissue lysates by a standard protocol. The batchwise purification scheme is well suited for multiple separations. The kit provides sufficient reagents to perform enrichments from up to 17.5 mg crude protein. Features and Benefits:
- Based on novel resin for sample enrichment for functional active protein kinases and other ATP-binding proteins
- Contains optimized buffers for maximum active protein recovery
- Compatible with 1D and 2D gel electrophoresis, LC/MS, and MALDI-TOF-MS
- Compatible with protein tyrosine kinase activity assays
Components:
| • | 100 mg | ATP-Binders Resin
| | • | 1 | PBS Tablet
| | • | 2 × 5 ml | 100 mM EDTA
| | • | 1.6 ml | 1 M DTT
| | • | 2 ml | ATP-Binders 6X Resin Conditioning Buffer
| | • | 7 ml | ATP-Binders 5X Bind Buffer
| | • | 5 ml | ATP-Binders Wash Buffer Concentrate
| | • | 1 ml | ATP-Binders Elution Buffer Concentrate
| | • | 5 ml | ATP-Binders 10X Nucleotide Mix
| | | (10 mM ADP, 10 mM AMP, 10 mM NADH, pH 7.0)
| | • | 1 ml | Protease Inhibitor Cocktail Set V, EDTA-Free, Lyophilized
| | • | 2 × 250 µl | 100 mM DTT
| | • | 100 µl | Activated Sodium Vanadate (200 mM Na3VO4, pH 10.0)
| | • | pkg/10 | Spin Filter, 2-ml capacity |
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