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NAD/NADH-Glo™ and NADP/NADPH-Glo™ Assays

Bioluminescent Assays to Detect NAD(P)+ and NAD(P)H Levels in Cells

  • Homogeneous, single-reagent-addition assays
  • Easily adapted for high throughput screening formats

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Catalog number selected: G9071

$ 499.00
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NAD/NADH-Glo™ and NADP/NADPH-Glo™ Assays
NAD/NADH/10ml
$ 499.00
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NADP_NADH_Background-Image

NAD/NADH-Glo™ Assay and NADP/NADPH-Glo™ Assays

The NAD/NADH-Glo and NADP/NADPH-Glo Assays are bioluminescent, homogeneous, single-reagent-addition methods to rapidly detect NAD+ and NADH and NADP+ and NADPH levels in cells and enzymatic reactions. Current methods to detect nicotinamide adenine dinucleotides lack sensitivity and reproducibility and require sample manipulation and large numbers of cells for cell-based applications. The assay is easily adapted for inhibitor screening in high-throughput formats. The sensitivity and robustness of the assay chemistry allow fewer cells to be used to detect individual nucleotides directly in multiwell plates or analysis of enzymes with low activity or with low Km values.

Low nanomolar sensitivity allows direct in-well detection and minimizes the amount of cells or enzyme required

Assay NAD/NADH-Glo™ Assay NADP/NADPH-Glo™ Assay
Limit of Detection (LOD) 1nM (25fmol/25µl) 1nM (25fmol/25µl)
Linearity 1–500nM 1–500nM
Signal-to-background ratio (S/B max) ~250 ~250
Cells/well for total dinucleotides 500–25,000 500–12,000
Cells/well for individual dinucleotides* 2,000–100,000 2,000–100,000
*Starting amount of cells; 1/4 of sample can be used to detect individual nucleotides
NAD_NADH_Foreground-Image

Assay Chemistry

In the presence of NADH, a reductase provided with the assay, reduces the proluciferin reductase substrate to form luciferin. Luciferin is quantified using Ultra-Glo™ Recombinant Luciferase, which is a component of the Luciferin Detection Reagent, and the light signal produced is proportional to the amount of NAD+ or NADH in the sample.

11738MC-W

Assays are Specific and Selective

Specificity is imparted to the NAD/NADH-Glo™ and NADP/NADPH-Glo™ Assay reactions by including NAD or NADP Cycling Enzyme and NAD or NADP Cycling Substrate to selectively measure nonphosphorylated and phosphorylated nucleotides, respectively. Accessory protocols are provided to allow separate measurements of NAD+ and NADH or NADP+ and NADPH and calculation of the ratio of the oxidized and reduced forms.

NADH, NADPH, NAD+ and NADP+ were prepared and diluted to the indicated concentrations in phosphate-buffered saline.

The NAD/NADH-Glo™ Assay is selective for NAD+ and NADH

The NADP/NADPH-Glo™ Assay is selective for NADP+ and NADPH

Detergent present in the reagent lyses cells, allowing detection of total cellular NAD+ and NADH or NADP+ and NADPH. For the NAD/NADH-Glo™ Assay, an accessory protocol is provided to allow separate measurements of NAD+ and NADH and calculation of the ratio of the oxidized and reduced forms. All reactions are performed directly in the sample wells.

Compatible with high throughput systems

The simple add-and-read protocols are compatible with automated and high-throughput workflow, and the flexible chemistries adapt to 96-, 384- and 1,536-well plate formats and high throughput systems. The yield Z′ values >0.7. Assays were performed using 8μl of the indicated concentration of NADH or NADPH with 8μl of the appropriate Luciferin Detection Reagent in 384-well plates. S/B = signal-to-background ratio.

NAD/NADH-Glo™ Assay Results

NADP/NADPH-Glo™ Assay Results

Specifications

You are viewing: G9071 Change Configuration

What's in the box?

Item Part # SizeConcentration

Reductase

G884A 1 × 55μl6mg/ml

Reductase Substrate

G885A 1 × 55μl

NAD Cycling Enzyme (lyophilized)

G887A 1 × 1 vial

NAD Cycling Substrate

G888A 1 × 1.25ml

Luciferin Detection Reagent

V859A 1 × 1 each

Reconstitution Buffer

V865A 1 × 10ml

SDS

Choose language:

Download SDSPDF (239 KB) – English (United States)

Certificate of Analysis

See all certificates for G9071

Use Restrictions

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

Storage Conditions

AA

Patents and Disclaimers

European Pat. No. 1131441 and Japanese Pat. No. 4520084.

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

Specifications

You are viewing: G9072 Change Configuration

What's in the box?

Item Part # SizeConcentration

Reductase

G884B 1 × 275μl6mg/ml

Reductase Substrate

G885B 1 × 275μl

NAD Cycling Enzyme (lyophilized)

G887A 1 × 1 vial

NAD Cycling Substrate

G888A 1 × 1.25ml

Luciferin Detection Reagent

V859B 1 × 1 each

Reconstitution Buffer

V865B 1 × 50ml

SDS

Choose language:

Download SDSPDF (239 KB) – English (United States)

Certificate of Analysis

See all certificates for G9072

Use Restrictions

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

Storage Conditions

AA

Patents and Disclaimers

European Pat. No. 1131441 and Japanese Pat. No. 4520084.

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

Specifications

You are viewing: G9081 Change Configuration

What's in the box?

Item Part # SizeConcentration

Reductase

G884A 1 × 55μl6mg/ml

Reductase Substrate

G885A 1 × 55μl

NADP Cycling Enzyme (lyophilized)

G889A 1 × 1 vial

NADP Cycling Substrate

G890A 1 × 275μl

Luciferin Detection Reagent

V859A 1 × 1 each

Reconstitution Buffer

V865A 1 × 10ml

SDS

Choose language:

Download SDSPDF (239 KB) – English (United States)

Certificate of Analysis

See all certificates for G9081

Use Restrictions

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

Storage Conditions

AA

Patents and Disclaimers

European Pat. No. 1131441 and Japanese Pat. No. 4520084.

Patent Pending.

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

Specifications

You are viewing: G9082 Change Configuration

What's in the box?

Item Part # SizeConcentration

Reductase

G884B 1 × 275μl6mg/ml

Reductase Substrate

G885B 1 × 275μl

NADP Cycling Enzyme (lyophilized)

G889A 1 × 1 vial

NADP Cycling Substrate

G890A 1 × 275μl

Luciferin Detection Reagent

V859B 1 × 1 each

Reconstitution Buffer

V865B 1 × 50ml

SDS

Choose language:

Download SDSPDF (239 KB) – English (United States)

Certificate of Analysis

See all certificates for G9082

Use Restrictions

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

Storage Conditions

AA

Patents and Disclaimers

European Pat. No. 1131441 and Japanese Pat. No. 4520084.

Patent Pending.

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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