Dehydrogenase-Glo™ Detection System

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Sensitive Detection of Dehydrogenase Enzyme Activity

  • Bioluminescent NAD(P)H detection toolkit that can be implemented rapidly to measure dehydrogenase activity in biological and biochemical samples
  • Extremely sensitive detection from just 10s of cells
  • Enables functional analysis of all major metabolic pathways

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Catalog number selected: J9010

$ 385.00
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Dehydrogenase-Glo™ Detection System
5ml
$ 385.00
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Simple Method to Measure Endogenous Dehydrogenase Activity

The Dehydrogenase-Glo™ Detection System provides a core bioluminescent NAD(P)H detection technology that can be used to measure dehydrogenase activity in cell lysates, tissue homogenates and other samples. The Dehydrogenase-Glo™ Detection System provides only the reagents required for bioluminescent NAD(P)H detection. To build a specific dehydrogenase assay, you will need to identify and source the dehydrogenase substrate. We provide straightforward instructions for assay optimization in the technical manual.

How the Dehydrogenase-Glo™ Detection System Works

Diagram illustrating how the Dehydrogenase-Glo™ Assay works.

The core technology provided in the Dehydrogenase-Glo™ Detection System measures NAD(P)H. To measure a dehydrogenase using this system, you need to identify and source an appropriate substrate. The endogenous dehydrogenase couples substrate oxidation to NAD(P)+ reduction. The reduced dinucleotides, NAD(P)H, are measured using a luciferase reaction. The amount of light produced is proportional to the NAD(P)H produced from specific dehydrogenase activity in the sample.

Functional Analysis of Metabolic Pathways Using Dehydrogenase-Glo™ Detection System

The Dehydrogenase-Glo™ Detection System can be used with commercially available substrates to characterize metabolic pathway activity.

Pathway

Dehydrogenase

Dehydrogenase Substrate

Dinucleotide (NAD/NADP)

Glycolysis

Glyceraldehyde 3-phosphate dehydrogenase (GAPDH)

d-glyceraldehyde 3-phosphate (Sigma Cat.# 39705)

NAD

Pentose Phosphate Pathway

Glucose 6-phosphate dehydrogenase (G6PDH)

Glucose 6-phosphate, (Sigma Cat.# 10127645001)

NADP

Pentose Phosphate Pathway

6-phosphogluconate dehydrogenase (6PGDH)

6-phosphogluconate (Sigma Cat.# P7877)

NADP

TCA Cycle

Isocitrate dehydrogenase (IDH)

d-isocitrate, (Sigma Cat.# 58790)

NADP

Measuring Dehydrogenase Activity in K562 Cells

Graph showing luminescence of dehydrogenases and controls.
Graph showing time-dependent GAPDH activity increase per cell density.
Graph showing specific activity of 4 dehydrogenases.

Measuring dehydrogenase activity in K562 cells. K562 cells were diluted twofold starting at 2.5 × 104 cells/ml in PBS, and 50μl was transferred into a 96-well assay plate. Dehydrogenase detection reagent was prepared for each dehydrogenase, and 50μl was added to sample wells. No-substrate controls (NAD only and NADP only) were included to determine background from nonspecific enzyme activity in K562 cells. Panel A. Luminescence was measured for dehydrogenases and controls after 60 minutes at different cell densities. Panel B. Time-dependent increase in GAPDH activity was shown at different cell densities. Luminescence was read every 15 minutes for 120 minutes. Solid black lines indicate linear range for each cell number. Panel C. Specific activity for each dehydrogenase was calculated. Data represent the average of three replicates with error bars indicating one standard deviation.

Do you want to learn more about our cellular energy metabolism tools and the innovative technology that powers these assays?

Specifications

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What's in the box?

Item Part # Size Concentration

Reductase

G884A 1 × 55μl 6mg/ml

Reductase Substrate

G885A 1 × 55μl

Luciferin Detection Solution

J135A 1 × 5ml

NAD

J136B 1 × 275μl

NADP+

J154B 1 × 100μl

Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

AA

Patents and Disclaimers

U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.

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