Assay of creatine kinase in microtiter plates using thio-NAD to allow monitoring at 405 nM

Analytical Biochemistry
J R Florini


An assay system for creatine kinase using microtiter plates and a plate reader that records absorbancies at 405 nM has been devised. The system is an adaptation of well-established assays that couple creatine kinase with the reactions catalyzed by hexokinase and glucose-6-phosphate dehydrogenase (G6PDH), to give a measurable increase in reduced pyridine nucleotide quantitated by absorbance at 340 nM. Two features of this system are modified for reading at 405 nM: (i) The thioamido derivative of NAD is used because its reduced form exhibits a substantial increase in absorbance at 405 nM, the most commonly available wavelength on microplate readers; and (ii) glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides is used because it can reduce either NAD or NADP (unlike most other G6PDH enzymes, which require NADP), thus making it unnecessary to use the more expensive thio-NADP. The rate of thio-NAD reduction is linear with enzyme concentration and time over a 20-fold range of concentrations of purified creatine kinase, and the assay also works well with myogenic cells allowed to grow and differentiate in the 96-well plate in which the assay is performed. This system offers considerable savings in cells, time, and materia...Continue Reading


Apr 30, 2004·Clinica Chimica Acta; International Journal of Clinical Chemistry·Masanori SeimiyaFumio Nomura
Apr 1, 1997·Tissue & Cell·T L WoodsS J Jones
Aug 1, 1997·Comparative Biochemistry and Physiology. Part A, Physiology·Y YunN H Ferrin
Mar 20, 2003·Molecular and Cellular Endocrinology·C D GalvinC M Nolan
Jun 7, 2000·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·S G VellemanD C McFarland
Mar 20, 2003·Comparative Biochemistry and Physiology. Toxicology & Pharmacology : CBP·D C McFarlandJ E Pesall
Feb 9, 2005·Proceedings of the National Academy of Sciences of the United States of America·Kathryn R WagnerR E Allen
Jun 19, 2007·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Brenda C HeldSteven H Grossman
May 18, 2004·Archives of Insect Biochemistry and Physiology·Ashli BrownSteven H Grossman
Jan 21, 2006·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·T G McDaneldD E Moody
Mar 7, 1997·The Journal of Biological Chemistry·S A CoolicanJ R Florini
Dec 31, 2015·Molecular and Cellular Biochemistry·Rachel L Harding, S G Velleman

Related Concepts

Macro-Creatine Kinase
Chinchilla Rabbits

Trending Feeds


Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Systemic Juvenile Idiopathic Arthritis

Systemic juvenile idiopathic arthritis is a rare rheumatic disease that affects children. Symptoms include joint pain, as well as fevers and skin rashes. Here is the latest research on this disease.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Cell Atlas Along the Gut-Brain Axis

Profiling cells along the gut-brain axis at the single cell level will provide unique information for each cell type, a three-dimensional map of how cell types work together to form tissues, and insights into how changes in the map underlie health and disease of the GI system and its crosstalk with the brain. Disocver the latest research on single cell analysis of the gut-brain axis here.

Myocardial Stunning

Myocardial stunning is a mechanical dysfunction that persists after reperfusion of previously ischemic tissue in the absence of irreversible damage including myocardial necrosis. Here is the latest research.

Pontocerebellar Hypoplasia

Pontocerebellar hypoplasias are a group of neurodegenerative autosomal recessive disorders with prenatal onset, atrophy or hypoplasia of the cerebellum, hypoplasia of the ventral pons, microcephaly, variable neocortical atrophy and severe mental and motor impairments. Here is the latest research on pontocerebellar hypoplasia.

Central Pontine Myelinolysis

Central Pontine Myelinolysis is a neurologic disorder caused most frequently by rapid correction of hyponatremia and is characterized by demyelination that affects the central portion of the base of the pons. Here is the latest research on this disease.


Trichotillomania is an impulse control disorder characterized by the recurrent pulling of one's hair resulting in noticeable hair loss. Here are the latest discoveries in this field.

DDX3X Syndrome

DDX3X syndrome is caused by a spontaneous mutation at conception that primarily affects females due to its location on the X-chromosome. DDX3X syndrome has been linked to intellectual disabilities, seizures, autism, low muscle tone, brain abnormalities, and slower physical developments. Here is the latest research.