6 Phosphogluconate Assay Kit (Colorimetric) (ab211071)

Overview

  • Product name

    6 Phosphogluconate Assay Kit (Colorimetric)
  • Detection method

    Colorimetric
  • Sample type

    Cell Lysate, Tissue Lysate
  • Assay type

    Quantitative
  • Sensitivity

    20 µM
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    6-Phosphogluconate Assay Kit (Colorimetric) (ab211071) provides a convenient method to detect 6-phosphogluconate (6-PGA) in cell and tissue lysates. In this assay, 6-phosphogluconate is converted to ribulose-5-phosphate by 6-phosphogluconate dehydrogenase in the presence of NAD+. NADH formed in the reaction reduces the probe present in the reaction mix, generating a strong absorbance at OD 450 nm. This absorbance is proportional to the amount of 6-PGA present in the samples.This assay kit is fast, sensitive and high-through adaptable. It can measure less than 20 µM of 6-phosphogluconate present in the sample.

  • Notes

    6-phosphogluconate (6-phosphgluconic acid, 6-PGA) is an intermediate of both Pentose Phosphate Pathway (PPP) and Entner-Doudoroff Pathway. In the Pentose Phosphate Pathway, 6-PGA is converted by 6-phosphogluconate dehydrogenase into ribulose-5-phosphate, a precursor for the synthesis of nucleic acids. In prokaryotes, 6-PGA is the main metabolite of the Entner-Doudoroff pathway, and is converted into pyruvate by both 6-phosphogluconate dehydratase and 2-keto-3-Deoxyphosphogluconate aldolase.

    Recent studies show that long-term exposure to glucose perturbs the PPP, causing significant accumulation of 6-PGA and impairment of beta cell function in the pancreas. Measurement of 6-PGA levels is important when evaluating therapies for diabetes research.

    In pathological bacteria, measurement of 6-PGA levels and functionality of Entner-Doudoroff pathway can be used for clinical and diagnostic purposes.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    PGA Assay Buffer 1 x 25ml
    6-PGA Enzyme (15 mg) 1 vial
    6-PGA Standard (10 mg) 1 vial
    6-PGA Substrate Mix (25 mg) 1 vial

Images

  • Typical 6-phosphogluconate (6-PGA) standard calibration curve.

  • Measurement of 6-PGA in lysates from rat liver (160 µg), rat kidney (120 µg) and rat heart (60 µg).

Protocols

References

ab211071 has not yet been referenced specifically in any publications.

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