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Performance of Ad-A-Gene Cyclic AMP Response Element-Nitroreductase (CRE-NTR): an adenoviral vector gene delivery system

siently expressed within those target cells at basal levels. When activated, CREB binds to the CRE sequences within the CRE-NTR gene, leading to promoter-induced up-regulation of NTR production. NTR is detected using the pro-fluorescent substrate, CytoCy5S™ (9). Quantitation of the NTR signal indicates the strength of the cellular response to the physical or chemical stimuli being investigated.

The CRE-NTR gene reporter assay described here was performed in live- and fixed-cell format in microplates. Assay plates were imaged on IN Cell Analyzer 1000 and read on a Tecan™ Ultra fluorimeter.


Materials
Reagents and instrumentation
Ad-A-Gene CRE-NTR GDS 40001

IN Cell Analyzer 1000 25-8010-26

IN Cell Analyzer 1000 Seat License* 25-8098-22

Object Intensity Analysis Module
(for IN Analyzer Cell 1000)† 25-8010-56

Nuclear Trafficking Analysis Module
(for IN Cell Analyzer 1000)† 25-8010-31

CytoCy5S, 0.2 mg PA76140

* A seat license is a cost-effective single-user or server license that gives access to all ready-to-use Image Analysis Modules provided with IN Cell Analyzer. License holders have access to all appropriate analysis software and more licenses can be purchased as the number of users grows.

† Available to seat license holders only.


Additional materials required
Tecan Ultra fluorimeter (Tecan group Ltd)

Culture medium: McCoy’s 5A (Sigma) with 10% fetal bovine serum (F
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