Team:Debrecen-Hungary/protocols/ Enzymatic activity with Victor machine

Scientific Background

The Wallac 1420 VICTOR2 is a multilabel, multitask plate reader designed to support the future demands of industrial and academic laboratories for multiple assay technologies on a single platform. An extended version of the successful Wallac VICTOR multilabel reader, the VICTOR2 allows immediate access to more than 10 counting modes, covering all of the main nonradioactive counting technologies.

VICTOR2 accepts all types of microtitration plates with between 1 and 864 (1536 fluorescence and time-resolved fluorescence) wells, as well as Petri dishes, slides, filters and Terasaki plates. All models include scanning, shaking and kinetics modules. To support individual applications in cell or molecular biology, binding studies, environmental and food testing, toxicology and drug screening, VICTOR2 can also be supplied with dispenser module, temperature control, bottom reading, high density reading and various other options.

Overview

Materials

Procedure

1. Pepare the following approximately half an hour before you start the B-gal and the Luciferase measurements

Switch on the machine VICTOR

Turn on the computer

Log in with your username

Open the program WALLAC 1420 WORKSTATION

2. Open main controll panel

Open the TOOLS tab and choose EXPLORER, after it you choose the BRAZDA* map

Open the corresponding file to measure the B-gal absorbance

choose the number of wells

save settings

3. go back to the main controll panel and choose the file programm within the PROTOCOL tab

4. Put the plate into the machine in right direction

5. Start the measurement

you may check live the measured absorbance values, if you click to the LIVE DISPLAY tab

6. To measure Luciferase activity, follow steps 1-4 with the corresponding file. Before step 4. add the Luciferase substrate

7. Use the icon which is a part of the main control panel for Dispenser maintenance

8. Appoint fill with the volume 500μL

9. Start measurement You may check live the measured Luciferase values, if you click to the LIVE DISPLAY tab

10. After the measurements empty the machine from the substrate

11. To wash the machine with water have to appoint flush (and the volume 500μL after it click ok) and then empty again

11. Turn off the VICTOR and then the computer

Notes & troubleshooting

References

1. Ping Wua,1, Lei Zhu1, Ulf-Hkan Stenman1 and Jari Leinonen1 "Immunopeptidometric Assay for Enzymatically Active Prostate-Specific Antigen" Clinical Chemistry. 2004;50:125-129

2. Néstor A. Chamoles, , Gabriela Niizawa, Mariana Blanco, Daniela Gaggioli and Carina Casentini "Glycogen storage disease type II: enzymatic screening in dried blood spots on filter paper" Clinica Chimica Acta Volume 347, Issues 1-2, September 2004

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