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  • Luminescence stability improvement in liposome-based homogeneous luminescence resonance energy transfer.

Luminescence stability improvement in liposome-based homogeneous luminescence resonance energy transfer.

Analytical biochemistry (2013-05-25)
Sari Pihlasalo, Jenny Granberg, Pekka Hänninen, Jouko Peltonen, Harri Härmä
ABSTRACT

A stable liposome-based time-resolved luminescence resonance energy transfer (TR-LRET) assay was developed based on the interaction of biotinylated lipids and streptavidin. Eu(3+) ion chelated to 4,4,4-trifluoro-1-(2-naphthalenyl)-1,3-butanedione and trioctylphosphine oxide was incorporated into liposomes. Acceptor-labeled streptavidin bound to biotinylated lipids of the liposomes enables TR-LRET. A stable assay performance was achieved by optimization. High Eu(3+) signal and stability, low variation, and sensitivity below 100 pM for free biotin was achieved by incorporating the chelate into liposomes containing cholesterol in a carbonate buffer. Potentially, the stable assay compared with the assay without cholesterol offers an improved platform to liposome-based detection systems.

MATERIALS
Product Number
Brand
Product Description

Supelco
Biotin, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
Biotin, tested according to Ph. Eur.
Sigma-Aldrich
Biotin, ≥99.0% (T)
Sigma-Aldrich
Biotin, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
Biotin, meets USP testing specifications
Biotin, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Biotin, Vetec, reagent grade, ≥99%
Supelco
Biotin, Pharmaceutical Secondary Standard; Certified Reference Material