Glucose Uptake Assay试剂盒(Colorimetric)

产品详情

How the assay works

Glucose Uptake Assay Kit (Colorimetric) is an assay that uses the glucose analog 2-deoxyglucose (2-DG) to detect and quantify glycose uptake in cells. 2-deoxyglucose (2-DG) is used in glucose uptake assay protocol 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 37°C
- Wash cells with PBS to remove exogenous 2-DG
- Lyse cells with extraction buffer and repeated pipetting
- Freeze/thaw lysates and heat at 85°C 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 37°C
- Add extraction buffer and heat to 90°C 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 37°C

How other researchers are using Glucose Uptake Assay ab136955

Glucose Uptake Assay Kit (Fluorometric) has been used in a variety of sample types including:
Human cervical epithelium HeLa and Human neuroblastoma SH-SY5Y ¹
Human osteosarcoma cancer cell lines 143B and U2OS ²
Human breast cancer cell lines ³
References:

1-Son S et al. 2023
2-Mei Z al. 2024
3-Peng Z et al. 2024

Related and recommended products

For fluorometric detection, we recommend Glucose Uptake Assay Kit (Fluorometric) ab136956

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