Abstract
Nerve growth factor (NGF), a neurotrophin critical to neuronal viability, has become a popular research focus for the treatment of neurodegenerative diseases. Enzyme-linked immunosorbent assays (ELISA) allow the quantification of cellular proteins, such as NGF, secreted into conditioned culture media. However, off-the-shelf reagents and kit components may require optimization depending on the specific specimens and/or antibodies that will be utilized with the ELISA assay. Herein we describe a protocol for developing a sandwich ELISA, incorporating NGF-specific primary and secondary antibodies capable of detecting NGF at concentrations as low as 10 pg/mL.
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References
Thermo Scientific Pierce Assay Development Technical Handbook: Version 2 (2011) Thermo Fisher Scientific, Inc. https://tools.thermofisher.com/content/sfs/brochures/1602127-Assay-Development-Handbook.pdf. Accessed 26 Mar 2016
Schuurs AH, van Weemen BK (1980) Enzyme-immunoassay: a powerful analytical tool. J Immunoass 1(2):229–249. doi:10.1080/01971528008055786
Houlgatte R, Mallat M, Brachet P, Prochiantz A (1989) Secretion of nerve growth factor in cultures of glial cells and neurons derived from different regions of the mouse brain. J Neurosci Res 24(2):143–152. doi:10.1002/jnr.490240204
Yoon SO, Casaccia-Bonnefil P, Carter B, Chao MV (1998) Competitive signaling between TrkA and p75 nerve growth factor receptors determines cell survival. J Neurosci 18(9):3273–3281
Soligo M, Protto V, Florenzano F, Bracci-Laudiero L, De Benedetti F, Chiaretti A, Manni L (2015) The mature/pro nerve growth factor ratio is decreased in the brain of diabetic rats: analysis by ELISA methods. Brain Res 1624:455–468. doi:10.1016/j.brainres.2015.08.005
Davis JB (2016) Developing an Alzheimer's disease therapeutic: identification and validation of signaling pathways involved in small molecule induction of nerve growth factor. Ph.D. diss., George Mason University, Ann Arbor, MI
Comparison of ABTS, TMB, and OPD Peroxidase Substrate Systems. Kirkegaard & Perry Laboratories, Inc. https://www.kpl.com/docs/techdocs/ML108.PDF. Accessed 26 March 2016 2016
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Davis, J.B. (2017). ELISA for Monitoring Nerve Growth Factor. In: Espina, V. (eds) Molecular Profiling. Methods in Molecular Biology, vol 1606. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6990-6_10
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DOI: https://doi.org/10.1007/978-1-4939-6990-6_10
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-6990-6
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