Cell Signaling Beta-Glo® Assay System Product Beta-Glo® Assay System For Research Use Only. Not for Use in Diagnostic Procedures. Description: The Beta-Glo® Assay System is a homogeneous method of quantitating β-galactosidase expression in mammalian cells. The system provides a bright luminescent signal that is stable over several hours in commonly used cell culture medium without prior sample processing. The homogeneous assay procedure involves the addition of a single reagent directly to cells cultured in serum-supplemented medium. Throughput rates of several thousand samples per hour may be achieved with high reproducibility under standard laboratory conditions. Features: • Bright Luminescent Signal: Quantitate with confidence using lowvolume formats or in samples with low-level expression. • Homogeneous Format: Perform fewer steps. Add a single reagent directly to cells in growth medium. • Stable Signal: Obtain flexibility and convenience when processing multiple plates. • Convenient: Achieve optimal assay performance at room temperature. • Flexible: Read the luminescent signal using any luminometer. Injectors are not required. • Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/ Storage Conditions: Store at –20°C. Protocol Beta-Glo® Assay System Technical Manual 0 1 2 3 4 5 6 Part# TM239 Size Cat.# Price (Fr) 10 ml E4720 172.00 100 ml E4740 1204.00 10 × 100 ml E4780 Pls. Enq. CAT Reporter Systems CAT Enzyme Assay System Product CAT Enzyme Assay System Available Separately Chloramphenicol Acetyltransferase n-Butyryl CoA Reporter Lysis 5X Buffer Size Conc. Size Cat.# Price (Fr) 50 reactions E1000 257.00 Cat.# Price (Fr) 100 u 10–14 u/µl E1051 96.00 255 µl 5 mg/ml 30 ml E1061 97.00 E3971 95.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: The CAT Enzyme Assay System offers two alternative methods for monitoring CAT enzyme activity in transfected cells: liquid scintillation counting (LSC) and thin layer chromatography (TLC). Either the LSC or TLC assays can be performed using the same cell extract. The TLC-based assay is less sensitive and more time-consuming to perform than the LSC assay but is useful as a visual confirmation of assay results. The resolved TLC reaction products are detected by autoradiography or phosphorimaging analysis. Chloramphenicol Acetyltransferase (CAT), encoded by a bacterial drugresistance gene, catalyzes the transfer of an acetyl group from acetyl-CoA to the 3´-hydroxy position of chloramphenicol. The enzyme is suitable as a standard in CAT assays of crude cell extracts. One unit is defined as the amount of enzyme required to transfer 1nmol of butyrate or acetate to chloramphenicol in one minute at 37°C. n-Butyryl CoA is suitable for use in the chloramphenicol acetyltransferase (CAT) reaction. Transfer of the n-butyryl moiety to chloramphenicol by the CAT enzyme allows enzyme activity to be monitored using liquid scintillation counting or thin layer chromatography formats. Features: • Fast: The assay is performed in as little as 2–3 hours. • Linear: The LSC assay is linear for three orders of magnitude of enzyme activity. • Sensitive: As little as 3 × 10–4 units (2pg) of CAT can be detected. • Robust: Reporter Lysis Buffer allows luciferase, CAT and β-galactosidase assays to be performed from the same cell extract. Storage Conditions: Reporter Lysis 5X Buffer may be stored at room temperature. Store other system components at –20°C. Protocol CAT Enzyme Assay System with Reporter Lysis Buffer Technical Bulletin Control Protein 1Protein 2 Beta-galactosidase activity determined using the Beta-Glo® Assay System with a yeast two-hybrid system. Image kindly provided by Dr. Brad Hook, Ph.D., University of Wisconsin, Madison. Part# TB084 15 For complete and up-to-date product information visit: www.promega.com/catalog 269 Reporter Assays and Transfection Log Luminescence (RLU) 6042MA Pagina 272
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