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2012 Promega catalogue

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Cell Signaling<br />

Beta-Glo ® Assay System<br />

Product Size Cat.# Price ($)<br />

Beta-Glo ® Assay System 10 ml E4720 154.00<br />

For Research Use Only. Not for Use in Diagnostic Procedures.<br />

100 ml E4740 1062.00<br />

10 × 100 ml E4780 Pls. Enq.<br />

Description: The Beta-Glo ® Assay System is a homogeneous method of<br />

quantitating β-galactosidase expression in mammalian cells. The system<br />

provides a bright luminescent signal that is stable over several hours in<br />

commonly used cell culture medium without prior sample processing. The<br />

homogeneous assay procedure involves the addition of a single reagent directly<br />

to cells cultured in serum-supplemented medium. Throughput rates of several<br />

thousand samples per hour may be achieved with high reproducibility under<br />

standard laboratory conditions.<br />

Features:<br />

• Bright Luminescent Signal: Quantitate with confidence using lowvolume<br />

formats or in samples with low-level expression.<br />

• Homogeneous Format: Perform fewer steps. Add a single reagent<br />

directly to cells in growth medium.<br />

• Stable Signal: Obtain flexibility and convenience when processing multiple<br />

plates.<br />

• Convenient: Achieve optimal assay performance at room temperature.<br />

• Flexible: Read the luminescent signal using any luminometer. Injectors are<br />

not required.<br />

• Choose Your Configuration: Learn more about our custom options for<br />

this product at: www.promega.com/myway/<br />

Storage Conditions: Store at –20°C.<br />

Protocol Part#<br />

Beta-Glo ® Assay System Technical Manual<br />

TM239<br />

Log Luminescence (RLU)<br />

6<br />

5<br />

4<br />

3<br />

2<br />

1<br />

0<br />

Control Protein 1 Protein 2<br />

Beta-galactosidase activity determined using the Beta-Glo ® Assay<br />

System with a yeast two-hybrid system. Image kindly provided by Dr.<br />

Brad Hook, Ph.D., University of Wisconsin, Madison.<br />

For complete and up-to-date product information visit: www.promega.com/catalog<br />

6042MA<br />

CAT Reporter Systems<br />

CAT Enzyme Assay System<br />

Product Size Cat.# Price ($)<br />

CAT Enzyme Assay System 50 reactions E1000 203.00<br />

Available Separately Size Conc. Cat.# Price ($)<br />

Chloramphenicol Acetyltransferase 100 u 10–14 u/µl E1051 82.00<br />

n-Butyryl CoA 255 µl 5 mg/ml E1061 103.00<br />

Reporter Lysis 5X Buffer 30 ml E3971 108.00<br />

For Research Use Only. Not for Use in Diagnostic Procedures.<br />

Description: The CAT Enzyme Assay System offers two alternative methods<br />

for monitoring CAT enzyme activity in transfected cells: liquid scintillation counting<br />

(LSC) and thin layer chromatography (TLC). Either the LSC or TLC assays<br />

can be performed using the same cell extract. The TLC-based assay is less<br />

sensitive and more time-consuming to perform than the LSC assay but is<br />

useful as a visual confirmation of assay results. The resolved TLC reaction<br />

products are detected by autoradiography or phosphorimaging analysis.<br />

Chloramphenicol Acetyltransferase (CAT), encoded by a bacterial drug-<br />

resistance gene, catalyzes the transfer of an acetyl group from acetyl-CoA to<br />

the 3´-hydroxy position of chloramphenicol. The enzyme is suitable as a<br />

standard in CAT assays of crude cell extracts. One unit is defined as the<br />

amount of enzyme required to transfer 1nmol of butyrate or acetate to<br />

chloramphenicol in one minute at 37°C.<br />

n-Butyryl CoA is suitable for use in the chloramphenicol acetyltransferase (CAT)<br />

reaction. Transfer of the n-butyryl moiety to chloramphenicol by the CAT<br />

enzyme allows enzyme activity to be monitored using liquid scintillation<br />

counting or thin layer chromatography formats.<br />

Features:<br />

• Fast: The assay is performed in as little as 2–3 hours.<br />

• Linear: The LSC assay is linear for three orders of magnitude of enzyme<br />

activity.<br />

• Sensitive: As little as 3 × 10 –4 units (2pg) of CAT can be detected.<br />

• Robust: Reporter Lysis Buffer allows luciferase, CAT and β-galactosidase<br />

assays to be performed from the same cell extract.<br />

Storage Conditions: Reporter Lysis 5X Buffer may be stored at room<br />

temperature. Store other system components at –20°C.<br />

Protocol Part#<br />

CAT Enzyme Assay System with Reporter Lysis Buffer Technical<br />

Bulletin<br />

TB084<br />

267<br />

15<br />

Reporter Assays and Transfection<br />

Section<br />

Contents<br />

Table of<br />

Contents

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