Luminescence - tecan infinite 200 Instructions For Use Manual

2. General Description
2.2.3

Luminescence

STOP
Switch on the instrument at least 15 minutes before starting a luminescence
Glow Type Chemi- or Bioluminescence
Bioluminescence Resonance Energy Transfer (BRET
Flash Luminescence
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For Flash reactions with the
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measurement. Some components need to warm up to guarantee stable
conditions for the measurement.
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The provides measurement of glow type chemi- or bioluminescence.
infinite 200
Glow type means that the luminescence assay glows much longer than a minute.
Luminescence substrates are available, which provide stable enough light output
over hours.
As an example, luminescence can be measured to determine the activity of an
enzyme labeled compound (-peroxidase, -phosphatase). Light emission results
from a luminescence substrate being decomposed by the enzyme. Under excess
of substrate the luminescence signal can be assumed to be proportional to the
abundance of the enzyme labeled compound. As with enzyme-based assays,
control of environmental conditions is rather critical (temperature, pH-value).
For practical aspects of luminescence assays see the following example:
Bioluminescence Methods and Protocols, ed. R.A. LaRossa, Methods in
Molecular Biology 102, Humana Press, 1998
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BRET is an advanced, non-destructive, cell-based assay technology that is
perfectly suited for proteomics applications, including receptor research and the
mapping of signal transduction pathways. BRET
between fusion proteins containing Renilla luciferase (Rluc) and a mutant of the
Green Fluorescent Protein (GFP). The BRET
oxidation of p.a. DeepBlueC™, a coelenterazine derivative that maximizes
spectral resolution for superior sensitivity. This homogeneous assay technology
provides a simple, robust and versatile platform with applications in basic
academic as well as applied research.
In flash type luminescence assays the measurement is only done during the
dispensing of the activating reagent or after a short delay time.
Over the past years luminescence substrates have been improved towards
providing more stable signals. In so-called glow type luminescence assays the
luminescence signal is spread over a wide time scale (e.g. a half-life of 30 min.)
200, see also 2.3.1 Measurement with Injectors.
infinite
Instructions for Use for infinite® 200 No. 30017581 Rev. No. 1.4
Caution
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)
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is based on energy transfer
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signal is generated by the
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2008-07