A rapid β-NADH-linked fluorescence assay for lactate dehydrogenase in cellular death

Jeffery H. Moran; Rick G. Schnellmann

doi:10.1016/1056-8719(96)00071-8

A rapid β-NADH-linked fluorescence assay for lactate dehydrogenase in cellular death

Jeffery H. Moran, Rick G. Schnellmann

Research output: Contribution to journal › Article › peer-review

62 Scopus citations

Abstract

Lactate dehydrogenase (LDH) release is a common marker of cellular death. Traditionally, the fraction of LDH released has been measured using a NADH-linked W-Vis spectrophotometric method. The limitation of this method is that samples are usually run serially and thus is time intensive. Therefore, we developed a NADH-linked LDH assay using a fluorescence plate reader that had a correlation of 0.95 with the traditional UV-Vis spectrophotometric method. Using rabbit renal proximal tubule suspensions at a concentration of 1 mg cellular protein/ml of media, the fluorescence assay can determine LDH release in 22 samples in 2 min using 12 μL of cellular homogenates and 150 μL of media. The parallel processing of samples and smaller volumes used in the fluorescence assay results in decreased analysis time and costs.

Original language	English (US)
Pages (from-to)	41-44
Number of pages	4
Journal	Journal of Pharmacological and Toxicological Methods
Volume	36
Issue number	1
DOIs	https://doi.org/10.1016/1056-8719(96)00071-8
State	Published - Sep 1996
Externally published	Yes

Keywords

Apoptosis
Cell death
Fluorescence assay
Lactate dehydrogenase
Oncosis
Phrases
Renal cells

ASJC Scopus subject areas

Toxicology
Pharmacology

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10.1016/1056-8719(96)00071-8

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title = "A rapid β-NADH-linked fluorescence assay for lactate dehydrogenase in cellular death",

abstract = "Lactate dehydrogenase (LDH) release is a common marker of cellular death. Traditionally, the fraction of LDH released has been measured using a NADH-linked W-Vis spectrophotometric method. The limitation of this method is that samples are usually run serially and thus is time intensive. Therefore, we developed a NADH-linked LDH assay using a fluorescence plate reader that had a correlation of 0.95 with the traditional UV-Vis spectrophotometric method. Using rabbit renal proximal tubule suspensions at a concentration of 1 mg cellular protein/ml of media, the fluorescence assay can determine LDH release in 22 samples in 2 min using 12 μL of cellular homogenates and 150 μL of media. The parallel processing of samples and smaller volumes used in the fluorescence assay results in decreased analysis time and costs.",

keywords = "Apoptosis, Cell death, Fluorescence assay, Lactate dehydrogenase, Oncosis, Phrases, Renal cells",

author = "Moran, {Jeffery H.} and Schnellmann, {Rick G.}",

note = "Funding Information: The authors would like to thank Dr. Grazyna Nowak and Ms. Shayla Waters for their review of the manuscript. This work was supported by Grant ES-04410 from the National Institute of Environmental Health Science.",

year = "1996",

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T1 - A rapid β-NADH-linked fluorescence assay for lactate dehydrogenase in cellular death

AU - Moran, Jeffery H.

AU - Schnellmann, Rick G.

N1 - Funding Information: The authors would like to thank Dr. Grazyna Nowak and Ms. Shayla Waters for their review of the manuscript. This work was supported by Grant ES-04410 from the National Institute of Environmental Health Science.

PY - 1996/9

Y1 - 1996/9

N2 - Lactate dehydrogenase (LDH) release is a common marker of cellular death. Traditionally, the fraction of LDH released has been measured using a NADH-linked W-Vis spectrophotometric method. The limitation of this method is that samples are usually run serially and thus is time intensive. Therefore, we developed a NADH-linked LDH assay using a fluorescence plate reader that had a correlation of 0.95 with the traditional UV-Vis spectrophotometric method. Using rabbit renal proximal tubule suspensions at a concentration of 1 mg cellular protein/ml of media, the fluorescence assay can determine LDH release in 22 samples in 2 min using 12 μL of cellular homogenates and 150 μL of media. The parallel processing of samples and smaller volumes used in the fluorescence assay results in decreased analysis time and costs.

AB - Lactate dehydrogenase (LDH) release is a common marker of cellular death. Traditionally, the fraction of LDH released has been measured using a NADH-linked W-Vis spectrophotometric method. The limitation of this method is that samples are usually run serially and thus is time intensive. Therefore, we developed a NADH-linked LDH assay using a fluorescence plate reader that had a correlation of 0.95 with the traditional UV-Vis spectrophotometric method. Using rabbit renal proximal tubule suspensions at a concentration of 1 mg cellular protein/ml of media, the fluorescence assay can determine LDH release in 22 samples in 2 min using 12 μL of cellular homogenates and 150 μL of media. The parallel processing of samples and smaller volumes used in the fluorescence assay results in decreased analysis time and costs.

KW - Apoptosis

KW - Cell death

KW - Fluorescence assay

KW - Lactate dehydrogenase

KW - Oncosis

KW - Phrases

KW - Renal cells

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ER -

A rapid β-NADH-linked fluorescence assay for lactate dehydrogenase in cellular death

Abstract

Keywords

ASJC Scopus subject areas

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