Jul 02, 2025
In vitro Generation and Quantification of AGEs
- Thien Ngoc Le1,
- Thien Ngoc Le1
- 1Flinders University
- Flinders University
Protocol Citation: Thien Ngoc Le, Thien Ngoc Le 2025. In vitro Generation and Quantification of AGEs . protocols.io https://dx.doi.org/10.17504/protocols.io.yxmvmbow5g3p/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working well.
Created: July 02, 2025
Last Modified: July 02, 2025
Protocol Integer ID: 221460
Keywords: quantification of age, age, generation, quantification
Abstract
A seven-step protocol provides how to generate and quantify AGEs in vitro.
Troubleshooting
In vitro Generation and Quantification of AGEs
To generate Advanced Glycation End Products (AGEs), 10 mL of human serum is incubated with 900 mg of D-glucose (Merck, Melbourne, Australia) at 37 °C in 5% CO2 atmosphere for 4–6 weeks.
The AGEs are dialyzed using Slide-A-Lyzer G3 dialysis cassettes (ThermoFisher Scientific, Melbourne, Australia) (10K molecular weight cut-off capacity) that allow de-glucose in the samples.
Sample (3 mL) is inserted in a lyzer device and stored in PBS buffer with a pH of 7.4 at 4 °C.
The buffer exchanges are performed every 24 hours.
Final sample of AGEs is collected after 3-day dialysis.
Sample of AGEs is stored in a freezer at -20 °C.
The concentration of AGEs in the human serum samples and control (commercial AGE-BSA) is measured using the AGE Assay Kit (ab238539; Abcam,Cambridge, UK), following the manufacturer's instructions.