Detection method Colorimetric (OD570-573nm)
Assay type Quantitative
Samples type Whole blood
Species All
Sample volume 120 μL (with K2 EDTA or heparin)
Metabolites measured NADP+, NADPH
Detection Limit 400 nM for NADP+ and 300 nM NADPH in the whole blood
Assay Length < 1h
Size 192 reactions
Number of samples 40 (measured in duplicates)
Storage Store reagents at -80°C
Intended use RUO
Shelf Life 6 months

Key Features

 

  • Highly accurate and sensitive.
  • Easy to handle. One step metabolite extraction is followed by separate measurement of NADP+ and NADPH in the extract. Detection time is less than 10 minutes.
  • High throughput. Can be easily adopted into high-throughput 96-well plate assay to analyze hundreds of samples per day.

Principals

The principle of the assay is a cyclic enzymatic reaction with a colorimetric end-point detection. First, NADP+ and NADPH metabolites are extracted together from a blood sample in a single step. Then the extract is divided into two parts revealing both NAPD+ and NADPH concentrations by removing one metabolite and stabilizing the other.

Reagents Provided   

Extraction BUFFER A, NAPD+ stabilizing reagent, NADPH stabilizing reagent, NADP+ standard stock, NADPH standard stock, Assay BUFFER C, Assay color reagent, Enzyme, Stop solution, Positive control

Kit Requires 

Multi-channel pipets. Two clear-bottom 96-well plates suitable for colorimetric assays, heat block with adjustable temperature (up to 80°C), spectrophotometric plate reader

Precautions and Warnings

For research use only.
The Stop Solution may cause skin, eye and respiratory irritation. Avoid breathing in fumes.
Assay color reagent may cause skin irritation. Handle with care, use gloves.
BUFFER A can cause eye irritation. Handle with care, use googles.
Read more in Q-NAD Safety Data Sheet (SDS)

General Information

Proprietary name: Q-NADP Blood: quantitative assay kit for NADP+ and NADPH in blood

More details how to prepare samples in sample preparation document.