Utilization of in situ ELISA method for examining Trk receptor phosphorylation in cultured cells

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Standard

Utilization of in situ ELISA method for examining Trk receptor phosphorylation in cultured cells. / Antila, Hanna; Autio, Henri; Turunen, Laura; Harju, Kirsi; Tammela, Päivi; Wennerberg, Krister; Yli-Kauhaluoma, Jari; Huttunen, Henri J; Castrén, Eero; Rantamäki, Tomi.

In: Journal of Neuroscience Methods, Vol. 222, 30.01.2014, p. 142-6.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Antila, H, Autio, H, Turunen, L, Harju, K, Tammela, P, Wennerberg, K, Yli-Kauhaluoma, J, Huttunen, HJ, Castrén, E & Rantamäki, T 2014, 'Utilization of in situ ELISA method for examining Trk receptor phosphorylation in cultured cells', Journal of Neuroscience Methods, vol. 222, pp. 142-6. https://doi.org/10.1016/j.jneumeth.2013.11.001

APA

Antila, H., Autio, H., Turunen, L., Harju, K., Tammela, P., Wennerberg, K., Yli-Kauhaluoma, J., Huttunen, H. J., Castrén, E., & Rantamäki, T. (2014). Utilization of in situ ELISA method for examining Trk receptor phosphorylation in cultured cells. Journal of Neuroscience Methods, 222, 142-6. https://doi.org/10.1016/j.jneumeth.2013.11.001

Vancouver

Antila H, Autio H, Turunen L, Harju K, Tammela P, Wennerberg K et al. Utilization of in situ ELISA method for examining Trk receptor phosphorylation in cultured cells. Journal of Neuroscience Methods. 2014 Jan 30;222:142-6. https://doi.org/10.1016/j.jneumeth.2013.11.001

Author

Antila, Hanna ; Autio, Henri ; Turunen, Laura ; Harju, Kirsi ; Tammela, Päivi ; Wennerberg, Krister ; Yli-Kauhaluoma, Jari ; Huttunen, Henri J ; Castrén, Eero ; Rantamäki, Tomi. / Utilization of in situ ELISA method for examining Trk receptor phosphorylation in cultured cells. In: Journal of Neuroscience Methods. 2014 ; Vol. 222. pp. 142-6.

Bibtex

@article{9a8bf74f62934e2db762c55eb3a8d564,
title = "Utilization of in situ ELISA method for examining Trk receptor phosphorylation in cultured cells",
abstract = "BACKGROUND: Trk receptor tyrosine kinases regulate multiple important neuronal processes during the development and in the adulthood. Tyrosine phosphorylation of Trk serves as the initial step in the Trk signaling pathway and indicates receptor' autocatalytic activity. However, methods allowing simple and large-scale Trk phosphorylation analyses in cultured cells are lacking.NEW METHOD: We describe an in situ phospho-Trk ELISA (enzyme-linked immunosorbent assay) method where cell culture, receptor stimulation and Trk phosphorylation analysis are all performed on the same multiwell plate.RESULTS: In situ phospho-Trk ELISA readily and specifically detects neurotrophin-induced Trk phosphorylation in cultured cells. A proof-of-concept small molecule screening of a library composed of 2000 approved drugs and other bioactive compounds was carried out using this novel method.COMPARISON WITH EXISTING METHODS: In situ phospho-Trk ELISA utilizes the principles and advantages of conventional sandwich ELISA in an in situ context.CONCLUSIONS: We describe a novel method that can be efficiently used to examine Trk receptor phosphorylation in cultured cells. Principally similar methods can be developed to examine the levels and signaling of any intracellular protein.",
keywords = "Animals, Cell Culture Techniques/instrumentation, Cell Line, Cells, Cultured, Cerebral Cortex/metabolism, Enzyme-Linked Immunosorbent Assay/methods, Hippocampus/metabolism, Mice, Mice, Transgenic, Nerve Growth Factors/metabolism, Neurons/metabolism, Phosphorylation, Rats, Receptor Protein-Tyrosine Kinases/metabolism, Receptor, trkB/genetics",
author = "Hanna Antila and Henri Autio and Laura Turunen and Kirsi Harju and P{\"a}ivi Tammela and Krister Wennerberg and Jari Yli-Kauhaluoma and Huttunen, {Henri J} and Eero Castr{\'e}n and Tomi Rantam{\"a}ki",
note = "Copyright {\textcopyright} 2013 Elsevier B.V. All rights reserved.",
year = "2014",
month = jan,
day = "30",
doi = "10.1016/j.jneumeth.2013.11.001",
language = "English",
volume = "222",
pages = "142--6",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Utilization of in situ ELISA method for examining Trk receptor phosphorylation in cultured cells

AU - Antila, Hanna

AU - Autio, Henri

AU - Turunen, Laura

AU - Harju, Kirsi

AU - Tammela, Päivi

AU - Wennerberg, Krister

AU - Yli-Kauhaluoma, Jari

AU - Huttunen, Henri J

AU - Castrén, Eero

AU - Rantamäki, Tomi

N1 - Copyright © 2013 Elsevier B.V. All rights reserved.

PY - 2014/1/30

Y1 - 2014/1/30

N2 - BACKGROUND: Trk receptor tyrosine kinases regulate multiple important neuronal processes during the development and in the adulthood. Tyrosine phosphorylation of Trk serves as the initial step in the Trk signaling pathway and indicates receptor' autocatalytic activity. However, methods allowing simple and large-scale Trk phosphorylation analyses in cultured cells are lacking.NEW METHOD: We describe an in situ phospho-Trk ELISA (enzyme-linked immunosorbent assay) method where cell culture, receptor stimulation and Trk phosphorylation analysis are all performed on the same multiwell plate.RESULTS: In situ phospho-Trk ELISA readily and specifically detects neurotrophin-induced Trk phosphorylation in cultured cells. A proof-of-concept small molecule screening of a library composed of 2000 approved drugs and other bioactive compounds was carried out using this novel method.COMPARISON WITH EXISTING METHODS: In situ phospho-Trk ELISA utilizes the principles and advantages of conventional sandwich ELISA in an in situ context.CONCLUSIONS: We describe a novel method that can be efficiently used to examine Trk receptor phosphorylation in cultured cells. Principally similar methods can be developed to examine the levels and signaling of any intracellular protein.

AB - BACKGROUND: Trk receptor tyrosine kinases regulate multiple important neuronal processes during the development and in the adulthood. Tyrosine phosphorylation of Trk serves as the initial step in the Trk signaling pathway and indicates receptor' autocatalytic activity. However, methods allowing simple and large-scale Trk phosphorylation analyses in cultured cells are lacking.NEW METHOD: We describe an in situ phospho-Trk ELISA (enzyme-linked immunosorbent assay) method where cell culture, receptor stimulation and Trk phosphorylation analysis are all performed on the same multiwell plate.RESULTS: In situ phospho-Trk ELISA readily and specifically detects neurotrophin-induced Trk phosphorylation in cultured cells. A proof-of-concept small molecule screening of a library composed of 2000 approved drugs and other bioactive compounds was carried out using this novel method.COMPARISON WITH EXISTING METHODS: In situ phospho-Trk ELISA utilizes the principles and advantages of conventional sandwich ELISA in an in situ context.CONCLUSIONS: We describe a novel method that can be efficiently used to examine Trk receptor phosphorylation in cultured cells. Principally similar methods can be developed to examine the levels and signaling of any intracellular protein.

KW - Animals

KW - Cell Culture Techniques/instrumentation

KW - Cell Line

KW - Cells, Cultured

KW - Cerebral Cortex/metabolism

KW - Enzyme-Linked Immunosorbent Assay/methods

KW - Hippocampus/metabolism

KW - Mice

KW - Mice, Transgenic

KW - Nerve Growth Factors/metabolism

KW - Neurons/metabolism

KW - Phosphorylation

KW - Rats

KW - Receptor Protein-Tyrosine Kinases/metabolism

KW - Receptor, trkB/genetics

U2 - 10.1016/j.jneumeth.2013.11.001

DO - 10.1016/j.jneumeth.2013.11.001

M3 - Journal article

C2 - 24239780

VL - 222

SP - 142

EP - 146

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

ER -

ID: 199431436