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Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR)

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskning

Standard

Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR). / Skeldal, Sune; Kjaergaard, Maj M; Alwasel, Saleh; Nyengaard, Jens R.

I: International Journal of Biochemistry and Molecular Biology, Bind 6, Nr. 2, 2015, s. 17-25.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskning

Harvard

Skeldal, S, Kjaergaard, MM, Alwasel, S & Nyengaard, JR 2015, 'Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR)', International Journal of Biochemistry and Molecular Biology, bind 6, nr. 2, s. 17-25.

APA

Skeldal, S., Kjaergaard, M. M., Alwasel, S., & Nyengaard, J. R. (2015). Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR). International Journal of Biochemistry and Molecular Biology, 6(2), 17-25.

CBE

Skeldal S, Kjaergaard MM, Alwasel S, Nyengaard JR. 2015. Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR). International Journal of Biochemistry and Molecular Biology. 6(2):17-25.

MLA

Vancouver

Skeldal S, Kjaergaard MM, Alwasel S, Nyengaard JR. Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR). International Journal of Biochemistry and Molecular Biology. 2015;6(2):17-25.

Author

Skeldal, Sune ; Kjaergaard, Maj M ; Alwasel, Saleh ; Nyengaard, Jens R. / Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR). I: International Journal of Biochemistry and Molecular Biology. 2015 ; Bind 6, Nr. 2. s. 17-25.

Bibtex

@article{5e205b2dae9b4bdaa63cd3f8bfe86a65,
title = "Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR)",
abstract = "Whereas the proform of the nerve growth factor (proNGF) is crucial for eliminating superfluous cells during neuronal development it also promotes apoptosis following brain trauma and neuronal injury. The apoptotic signal is elicited upon formation of a trimeric receptor complex also containing the vps10p domain receptor sortilin and the neurotrophin receptor p75(NTR). However, proNGF-induced receptor complex formation has been difficult to directly assess other than by western blotting. We here describe a fluorescence resonance energy transfer (FRET) based fluorescence plate reader assay to monitor the interaction between fluorescently tagged sortilin and p75(NTR) in live cells. The method is based on a standard fluorescent plate reader found in many biochemical laboratories and the results are evaluated using a microscopy-based quantified sensitized acceptor emission FRET approach making use of a pair of FRET standard constructs. As a result, the effect of proNGF on the interaction between sortilin and p75(NTR) can be evaluated in live cells allowing for screening and selection of therapeutic compounds interfering with proNGF-induced cell death.",
author = "Sune Skeldal and Kjaergaard, {Maj M} and Saleh Alwasel and Nyengaard, {Jens R}",
year = "2015",
language = "English",
volume = "6",
pages = "17--25",
journal = "International Journal of Biochemistry and Molecular Biology",
issn = "2152-4114",
publisher = "E-Century Publishing Corporation",
number = "2",

}

RIS

TY - JOUR

T1 - Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR)

AU - Skeldal, Sune

AU - Kjaergaard, Maj M

AU - Alwasel, Saleh

AU - Nyengaard, Jens R

PY - 2015

Y1 - 2015

N2 - Whereas the proform of the nerve growth factor (proNGF) is crucial for eliminating superfluous cells during neuronal development it also promotes apoptosis following brain trauma and neuronal injury. The apoptotic signal is elicited upon formation of a trimeric receptor complex also containing the vps10p domain receptor sortilin and the neurotrophin receptor p75(NTR). However, proNGF-induced receptor complex formation has been difficult to directly assess other than by western blotting. We here describe a fluorescence resonance energy transfer (FRET) based fluorescence plate reader assay to monitor the interaction between fluorescently tagged sortilin and p75(NTR) in live cells. The method is based on a standard fluorescent plate reader found in many biochemical laboratories and the results are evaluated using a microscopy-based quantified sensitized acceptor emission FRET approach making use of a pair of FRET standard constructs. As a result, the effect of proNGF on the interaction between sortilin and p75(NTR) can be evaluated in live cells allowing for screening and selection of therapeutic compounds interfering with proNGF-induced cell death.

AB - Whereas the proform of the nerve growth factor (proNGF) is crucial for eliminating superfluous cells during neuronal development it also promotes apoptosis following brain trauma and neuronal injury. The apoptotic signal is elicited upon formation of a trimeric receptor complex also containing the vps10p domain receptor sortilin and the neurotrophin receptor p75(NTR). However, proNGF-induced receptor complex formation has been difficult to directly assess other than by western blotting. We here describe a fluorescence resonance energy transfer (FRET) based fluorescence plate reader assay to monitor the interaction between fluorescently tagged sortilin and p75(NTR) in live cells. The method is based on a standard fluorescent plate reader found in many biochemical laboratories and the results are evaluated using a microscopy-based quantified sensitized acceptor emission FRET approach making use of a pair of FRET standard constructs. As a result, the effect of proNGF on the interaction between sortilin and p75(NTR) can be evaluated in live cells allowing for screening and selection of therapeutic compounds interfering with proNGF-induced cell death.

M3 - Journal article

C2 - 26823987

VL - 6

SP - 17

EP - 25

JO - International Journal of Biochemistry and Molecular Biology

JF - International Journal of Biochemistry and Molecular Biology

SN - 2152-4114

IS - 2

ER -