Cell Signaling SignaTECT® Protein Kinase Assay Systems Product SignaTECT® cAMP-Dependent Protein Kinase (PKA) Assay System SignaTECT® Protein Kinase C (PKC) Assay System SignaTECT® Protein Tyrosine Kinase (PTK) Assay System SignaTECT® Calcium/Calmodulin-Dependent Protein Kinase (CaM KII) Assay System SignaTECT® DNA-Dependent Protein Kinase Assay System SignaTECT® cdc2 Protein Kinase Assay System Size Cat.# Price (Fr) 96 reactions V7480 679.00 96 reactions V7470 679.00 96 reactions V6480 742.00 96 reactions V8161 679.00 96 reactions V7870 679.00 96 reactions V6430 679.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: The SignaTECT® Protein Kinase Assay Systems contain the proprietary SAM2® Biotin Capture Membrane. The streptavidin-coated SAM2® Membranes possess high binding capacity and high specificity characteristics, which produce lower backgrounds and higher signal-to-noise ratios compared to the traditional P81 phosphocellulose method of capture and measurement. The perforated and numbered membrane allows researchers to measure from 1 up to 96 kinase reactions. The SAM2® Membrane format does not require as much “hands-on” manipulation as other methods used to measure kinase activity. Following the kinase reaction, samples are spotted onto the SAM2® Membrane, and a series of short wash steps are performed to remove nonspecific label. The process is complete in less than 1 hour. In addition, the nature of the SAM2® Membrane allows it to be used under a variety of buffer and reaction conditions (e.g., cell extracts), which many other methods do not allow. Last, the high binding capacity allows use of the SignaTECT® Systems for kinetic studies. Each SignaTect® system contains specific biotinylated peptide substrates for the appropriate kinase as well as the necessary reaction components. The researcher must supply [γ-32P]ATP. Storage Conditions: Store all SignaTECT® Systems except V7470 at –20°C. Store Cat.# V7470 at –70°C. Protocol SignaTECT® cAMP-Dependent Protein Kinase (PKA) Assay System Technical Bulletin SignaTECT® Calcium/Calmodulin-Dependent Protein Kinase Assay System Technical Bulletin SignaTECT® cdc2 Protein Kinase Assay System Technical Bulletin Part# TB241 SignaTECT® Protein Kinase C (PKC) Assay System Technical Bulletin TB242 SignaTECT® Protein Tyrosine Kinase Assay System Technical Bulletin TB211 TB279 SignaTECT® DNA-Dependent Protein Kinase Assay System Technical Bulletin TB250 TB227 Biotin–(C)6–XXXX(Y/S/T)XXXX Biotinylated Peptide Substrate Protein Kinase Sample + + Reaction Buffer + [γ-32P]ATP Combine peptide substrate, [γ-32P]ATP, buffer and sample. Incubate at 30°C for 2–10 minutes. Terminate the kinase reaction with Termination Buffer. Biotin–(C)6–XXXX(Y/S/T)XXXX PO4 Capture the biotinylated peptide substrate by spotting reactions on the Membrane. SAM2® Biotin Capture Membrane Components Reaction Biotin–(C)6–XXXX(Y/S/T)XXXX PO4 Wash Membrane to remove unbound reaction components. Quantitate • Scintillation counter • Phosphorimaging system • Autoradiography Schematic diagram of the SignaTECT® Protein Kinase Assay protocol. Protocol steps to prepare, run and analyze a specific protein kinase activity using any of the SignaTECT® Protein Kinase Assay Systems. 3 For complete and up-to-date product information visit: www.promega.com/catalog 63 Cell Signaling 1542MA05_7A Pagina 66
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