PMID: 7526085Jun 1, 1994Paper

Assay of neuronal nitric oxide synthase by HPLC determination of citrulline

Journal of Neuroscience Methods
M Carlberg

Abstract

Biological membranes from different tissue sources were incubated for nitric oxide synthase (NOS) activity under standard conditions at 20 degrees C and compared with rat cerebellar cytosol. NOS activity was monitored as the formation of L-citrulline from L-arginine. Samples were purified on Amprep CBA cation-exchange minicolumns prior to derivatization with o-phthaldialdehyde (OPA) and HPLC analysis. The OPA derivatives of L-citrulline and L-arginine eluted well separated within 15 min during isocratic elution at room temperature. A linear relation between peak height and quantity of L-citrulline was seen down to the detection limit at 0.1 pmol L-citrulline. Formation of L-citrulline was measurable in rat cerebellar cytosol as well as in preparations not previously assayed for NOS activity, including rat colon, cat oesophagus and crayfish (Pacifastacus leniusculus) brain. The method provides a sensitive and non-radioactive method for assaying NOS activity in small tissue samples and in tissues with low to moderate levels of NOS activity.

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Citations

Jan 22, 1998·The Annals of Thoracic Surgery·E E TsengW A Baumgartner
Apr 10, 1999·The Annals of Thoracic Surgery·E E TsengW A Baumgartner
Mar 15, 2003·Microbiology and Immunology·Chihomi KatoKazuko Saito
Sep 9, 2006·Neuroscience Letters·Natalia B Saulskaya, Nellia V Fofonova
Jan 1, 1995·Advances in Neuroimmunology·K U Johansson, M Carlberg
Jun 30, 2001·Brain Research·R J GerritsA S Greene
Mar 23, 1999·The Annals of Thoracic Surgery·E E TsengW A Baumgartner
Jun 15, 2016·Indian Journal of Pharmacology·Sadaf A Karbelkar, Anuradha S Majumdar
Aug 1, 1996·Hypertension·D L Mattson, T G Bellehumeur
Jun 17, 1998·Stroke; a Journal of Cerebral Circulation·H OkamotoA G Hudetz
Feb 4, 1998·Hypertension·D L MattsonA W Cowley
Dec 18, 2001·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Kirk P ConradJackie Novak
Jul 14, 2000·American Journal of Physiology. Endocrinology and Metabolism·R HenningssonI Lundquist
Jan 8, 1999·Journal of Periodontal Research·M MatejkaH Sinzinger
Jun 19, 2002·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Mátyás SzentiványiAllen W Cowley
Sep 2, 1998·The American Journal of Physiology·R Henningsson, I Lundquist
Oct 26, 2000·Journal of Chromatography. B, Biomedical Sciences and Applications·K IgarashiN Castagnoli

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