Science Catalog 2012 Life Cell Signaling PepTag® Non-Radioactive Protein Kinase Assays Product PepTag® Non-Radioactive PKC Assay PepTag® Non-Radioactive cAMP-Dependent Protein Kinase Assay Size Cat.# Price (Fr) 120 reactions V5330 607.00 120 reactions V5340 607.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: The PepTag® Non-Radioactive Protein Kinase Assay Systems provide a rapid, sensitive and non-radioactive method to detect either Protein Kinase C (PKC) or Protein Kinase A (PKA) activity. The PepTag® Assays use brightly colored, fluorescent peptide substrates that are highly specific for PKC (PepTag® C1 Peptide-PLSRTLSVAAK) and PKA (PepTag® A1 PeptideLRRASLG). Phosphorylation of the peptide alters the net charge from +1 to –1. This change in the net charge allows the phosphorylated and nonphosphorylated versions of the substrate to be rapidly separated on an agarose gel at neutral pH. Using fluorescent detection, less than 2ng of purified kinase can be detected in less than 2 hours. The PepTag® Non-Radioactive Protein Kinase Assay Systems can detect kinase activity in partially purified samples as well as purified preparations of enzymes, making it a good choice for the rapid screening of column fractions or the screening of kinase activators and inhibitors. In addition to the assay components, each system includes purified kinase for use as a positive control. Features: • Nonradioactive: The fluorescent tag on the peptide substrate facilitates quantitation of the phosphorylation reaction without the use of radioactivity. • Low Background: Phosphorylation of the colored peptide supplied with the system is used to measure kinase activity; therefore phosphorylation of other substrates occurring naturally in the sample does not add to the kinase activity measured. • Convenient: Measure the phosphorylated peptide using a densitometer, spectrophotometer, 96-well plate reader, or fluorometer. Storage Conditions: Store at –70°C. Protocol PepTag® Assay for Non-Radioactive Detection of Protein Kinase C or cAMP-Dependent Protein Kinase Technical Bulletin colored peptide substrate (+1 net charge) Incubate peptide with kinase-containing sample. P = nonphosphorylated (+1 net charge) phosphorylated (–1 net charge) Separate phosphorylated and nonphosphorylated forms by agarose gel electrophoresis. Net Charge of Peptide +1 0 –1 cathode Schematic diagram of PepTag® Non-Radioactive Protein Kinase Assay procedure. anode Peptide Species nonphosphorylated P = phosphorylated Part# TB132 DNA-Dependent Protein Kinase Product DNA-Dependent Protein Kinase For Research Use Only. Not for Use in Diagnostic Procedures. Description: DNA-Dependent Protein Kinase (DNA-PK) phosphorylates several DNA-binding substrates in vitro, including the tumor suppressor protein p53, the SV40 large T antigen and several transcription factors. DNA-PK is thought to play a role in controlling gene regulation and cell growth. DNA-PK is isolated from HeLa nuclear extracts as a complex consisting of a 400kDa catalytic subunit and a 155kDa heterodimeric DNA-binding component named Ku, which itself consists of subunits of approximately 85kDa and 70kDa. Storage Conditions: Store at –70°C. Protocol DNA-Dependent Protein Kinase Product Information cGMP-Dependent Protein Kinase Product cGMP-Dependent Protein Kinase (α-Isozyme) For Research Use Only. Not for Use in Diagnostic Procedures. Size Cat.# Price (Fr) 6,000 u V5171 364.00 Description: cGMP-Dependent Protein Kinase is a serine/threonine protein kinase present in smooth muscle and a variety of other tissues. The kinase is a 78kDa polypeptide composed of a regulatory domain and a catalytic domain and is active as a homodimer. Specific Activity: >1,000u/μg (kinase activity). Features: • Highly Pure: cGMP-Dependent Protein Kinase has been purified by the method of Corbin and Doskeland and is >90% pure as determined by SDS-PAGE (single band). Storage Conditions: Store at –70°C. Protocol cGMP-Dependent Protein Kinase (Alpha Isozyme) Product Information 64 Part# 9PIV517 For complete and up-to-date product information visit: www.promega.com/catalog Part# 9PIV581 Size Cat.# Price (Fr) 2,500 u V5811 461.00 Protein Kinases cAMP-Dependent Protein Kinase, Catalytic Subunit Product cAMP-Dependent Protein Kinase, Catalytic Subunit Size Conc. 2,500 u 1.5–3 mg/ml For Research Use Only. Not for Use in Diagnostic Procedures. Description: The purified 40kDa cAMP-Dependent Protein Kinase (PKA), Catalytic Subunit, may be used to phosphorylate target proteins or for in vitro enzymological studies of neural and hormonal signal transduction. Intracellular targets include ion channels, transcriptional activator proteins, and regulatory enzymes of glycogen metabolism. Features: • Highly Pure: The PKA Catalytic Subunit is purified from a recombinant E. coli strain expressing the catalytic subunit of bovine PKA and is 90% pure. Storage Conditions: Store at –70°C. Protocol cAMP-Dependent Protein Kinase, Catalytic Subunit Product Information Part# 9PIV516 Cat.# Price (Fr) V5161 331.00 0383MB Pagina 67
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