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Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol TRAP Assay
Published on: September 2, 2014
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Telomerase stability and evaluation of real-time telomeric repeat amplification protocol
Aleksandra R Vukašinović1, Jelena M Kotur-Stevuljević1, Vid Mlakar2
1a Faculty of Pharmacy, Department of Medical Biochemistry , University of Belgrade , Belgrade , Serbia.
Scandinavian Journal of Clinical and Laboratory Investigation
|February 16, 2019
Summary
Telomerase activity measurement requires specific sample storage conditions for reliability. Cell lines offer stable standards for this enzyme, a potential biomarker for various diseases.
Area of Science:
- Biochemistry
- Molecular Biology
- Biomarker Discovery
Background:
- Telomerase, an RNA-directed polymerase, functions as a reverse transcriptase.
- Telomerase is implicated in disease pathology and recognized as a potential biomarker.
Purpose of the Study:
- Determine optimal sample storage conditions and analysis timeframes for telomerase activity.
- Validate the reliability of the real-time telomeric repeat amplification protocol (TRAP) for enzyme activity measurement.
- Identify suitable standard samples for telomerase activity assays.
Main Methods:
- Peripheral blood leukocytes from healthy individuals and cancer patients were analyzed.
- Telomerase activity was measured using the real-time telomeric repeat amplification protocol (TRAP).
- Standard sample evaluation utilized nuclear magnetic resonance (NMR) spectroscopy.
Main Results:
- Samples from cancer patients showed preserved telomerase activity at -20°C for 14 days.
- Optimal storage for healthy individuals' samples was determined to be -80°C.
- Significant decreases in telomerase activity were observed after freeze-thaw cycles, particularly in healthy individuals' samples (cycle 3) and cancer patients' samples (cycle 5).
- Cell line samples demonstrated superior stability for storage and freeze-thaw cycles, serving as reliable standards confirmed by NMR.
Conclusions:
- Telomerase enzyme exhibits adequate stability for biomarker analysis.
- The TRAP method is reliable and reproducible for bio-analytical applications.
- Telomerase is a promising and reliable biomarker for disease detection and monitoring.
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