Screen Quest™ Colorimetric Glucose Uptake Assay Kit
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
Platform
Absorbance microplate reader
Absorbance | 570/610 nm |
Recommended plate | Clear bottom |
Components
Example protocol
AT A GLANCE
Protocol summary
- Plate cells and treat the cells as desired
- Add 10 µL/well 2-DG and incubate at 37oC for 20-40 minutes
- Wash cells and lyse cells
- Add 50 µL/well of 2-DG Uptake Assay working solution
- Incubate at RT for 30 to 120 minutes
- Monitor OD ratio increase at 570/610 nm
Important notes
Thaw all the kit components at room temperature before starting the experiment. The above mentioned protocol is for one 96-well plate.
PREPARATION OF STOCK SOLUTION
KRPH Buffer stock solution (1X):
Add 20 mL of KRPH Buffer (5X) (Component H) to 80 mL of deionized water and mix them well. Note: 50 mL volume of 1× KRPH Buffer is enough for approximately one 96-well plate. Prepare the needed volume proportionally. Store the unused 1×KRPH at 4oC or -20oC.
PREPARATION OF WORKING SOLUTION
1. NADP working solution:
Add 100 µL of H2O into the vial of NADP (Component G) and mix them well.
2. Enzyme Probe working solution:
Add 5 mL of Assay Buffer (Component F) into the bottle of Enzyme Probe (Component E) and mix them well.
3. 2-DG Uptake Assay working solution:
Add 100 µL of NADP working solution into the Enzyme Probe working solution and mix them well. Note: These quantities are good for one 96-well plate.
SAMPLE EXPERIMENTAL PROTOCOL
Important: This protocol can be used as guidelines to culture 3T3-L1 adipocytes for 2-DG uptake.
- Plate cells in growth medium at 50,000-80,000 cells/well/100 µL/96-well or 12,500-20,000 cells/well/25 µL/384-well black wall/clear bottom cell culture Poly-D lysine plate for 4-6 hours before experiment.
- Remove the cell plate from the incubator, aspirate the medium from the wells, and deprive the cells with 100 µL/well (96 well-plate) or 25 µL/well (384 well-plate) serum free medium. Incubate the cells at 37oC, 5% CO2 incubator for 6 hours to overnight.
- Remove the cell plate from the incubator, aspirate the medium from the wells, and gently wash the cells twice with 100 µL/well 1× KRPH buffer.
- Add 90 µL/well Glucose Uptake Buffer (Component B) and incubate the cells at 37oC, 5% CO2 incubator for 1 hour.
- Stimulate with or without insulin or compound of test for 20 min. Add 10 µL/well of the 10× insulin solution to a final concentration of 1 µM or 10× compound solution of test. And also add 10 µL insulin vehicle buffer or compound vehicle buffer to the untreated wells as control, and incubate at 37oC, 5% CO2 incubator for 20 mins.
- For glucose uptake inhibition study, add 10× Phloretin to a final concentration of 200 uM or inhibitors of test, and incubate at 37oC, 5% CO2 for 2-5 mins. Note: 10 mL inhibitor vehicle buffer is suggested to be added to both the insulin treated and untreated wells as control. Phloretin treated cells can be used as positive control.
- Add 10 µL/well 2-DG solution (Component A) to each well, and incubate at 37oC, 5% CO2 incubator for 20-40 mins. For negative controls, leave some wells untreated with insulin, inhibitor and 2-DG.
- After treatment, remove solution in each well and gently wash cells 3 times, 100 µL/well with KRPH Buffer to remove the extra 2-DG from the solution. Remove KRPH Buffer from the wells.
- Add 25 µL/well Acidic Lysis Buffer (Component C) to each well and incubate at 37oC for 20 min to lyse the cells. And the 2DG uptake assay mixture could be prepared in the meantime.
- Add 25 µL/well Neutralization Buffer (Component D) to each well, mix thoroughly, leave at room temperature for 5-10 minutes to neutralize the cell lysate.
- Add 50 µL of 2DG Uptake Assay working solution to each well of 2-DG6P standard (Not provided) or cell lysate.
- Incubate the reaction at room temperature for 30 minutes to 2 hours, protected from light.
- Monitor the absorbance ratio increase at 570/610 nm with an absorbance plate reader.
Images

Citations
Authors: Li, Zhuo and Jiang, Ying and Liu, Jian and Fu, Huifeng and Yang, Quan and Song, Wei and Li, Yuanwei
Journal: International Journal of Oncology (2023): 1--13
Authors: Xia, Peng and Zhang, Hao and Lu, Haofeng and Xu, Kequan and Jiang, Xiang and Jiang, Yuke and Gongye, Xiangdong and Chen, Zhang and Liu, Jie and Chen, Xi and others,
Journal: Cancer Communications (2023)
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Journal: The Kaohsiung Journal of Medical Sciences (2022): 1080--1092
Authors: Chen, Geng and Jiang, Jiayun and Wang, Xiaofei and Feng, Kai and Ma, Kuansheng
Journal: Computational and Mathematical Methods in Medicine (2021)
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Authors: Lu, Jie and Montgomery, Blake K and Chatain, Gr{\'e}goire P and Bugarini, Alejandro and Zhang, Qi and Wang, Xiang and Edwards, Nancy A and Ray-Chaudhury, Abhik and Merrill, Marsha J and Lonser, Russell R and others,
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