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Glucose Uptake Assay Kit (Colorimetric) (ab136955)

Key features and details

  • Assay type: Cell-based (quantitative)
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 3 hr
  • Sample type: Adherent cells, Suspension cells
  • Sensitivity: 0.01 nmol/well

Overview

  • Product name

    Glucose Uptake Assay Kit (Colorimetric)
    See all Glucose uptake kits

  • Detection method

    Colorimetric

  • Sample type

    Adherent cells, Suspension cells

  • Assay type

    Cell-based (quantitative)

  • Sensitivity

  • Assay time

    3h 00m

  • Species reactivity

    Reacts with: Mammals, Other species

  • Product overview

    Glucose Uptake Assay Kit (Colorimetric) (ab136955) is a highly sensitive and easy to use non-radioactive assay kit which can detect glucose uptake as low as 10 pmol/well in a variety of cell types.


    2-deoxyglucose (2-DG) is used in glucose uptake assay protocols because of its structural similarity to glucose. 2-DG is taken up by glucose transporters and metabolized to 2-DG-6-phosphate (2-DG6P). 2-DG6P cannot be further metabolized, and thus accumulates within cells. The accumulated 2-DG6P is directly proportional to 2-DG (or glucose) uptake by cells. In this assay, the 2-DG6P is oxidized to generate NADPH, the level of which can be determined by an enzymatic recycling amplification reaction.


    Glucose uptake assay protocol summary:
    - prepare cells with suitable glucose starvation / uptake stimulation depending on experimental set-up
    - add 2-DG to cells and incubate for 20 mins at 37ºC
    - wash cells with PBS to remove exogenous 2-DG
    - lyse cells with extraction buffer and repeated pipetting
    - freeze/thaw lysates and heat at 85ºC for 40 min
    - cool on ice for 5 min
    - add neutralizing buffer, spin and transfer supernatant to new tubes
    - add supernatants and standards to wells
    - add reaction mix A and incubate for 1 hr at 37ºC
    - add extraction buffer and heat to 90ºC for 40 min
    - cool on ice for 5 min and add neutralizing buffer
    - add reaction mix B
    - analyze every 2-3 mins on microplate reader in kinetic mode at 37ºC

  • Notes

    Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.

  • Platform

    Microplate reader

Properties

Properties

Images

  • Standard curve and example data.
    Standard curve and example data.
    2-DG6P Standard curve (a) and 2-DG uptake in 3T3-L1 cells (b), Human adipocytes (c) and HeLa cells (d) respectively. I=Insulin; P=Phloretin.
  • Functional Studies - Glucose Uptake Assay Kit (ab136955)
    Functional Studies - Glucose Uptake Assay Kit (ab136955)

    Glucose uptake in 3T3-L1 adipocytes stimulated with insulin (I). 3T3-L1 adipocytes were differentiated using:

     

    Dexamethasone ab120743 (1mM, 1:1000)

    IBMX ab120840 (11.5 mg/mL, 1:100)

    Insulin ab123768 (1 mg/mL, 1:1000)

  • Assay Procedure
    Assay Procedure
    Step A: 2-DG oxidation to generate NADPH; Step B: NADPH recycling amplification Reaction.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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