Product nameGlycated Protein Assay Kit
ab219802 enables the highly sensitive detection of glycated proteins (advanced glycation endproducts, AGEs) using a simple polyacrylamide gel electrophoresis (PAGE)-based method. This kit is compatible with a range of biological samples, including lysates, plasma, sera or tissue homogenates.
Glucose and other metabolites of glycolysis react directly with important cellular components such as DNA, lipids and protein through a process known as glycation. During glycation, reducing sugar molecules react with the amino groups of amino acids such as those found on lysine, arginine and protein N-termini, ultimately leading to the formation of complex and stable AGEs.
This kit uses Fluorescein-phenylboronate gel electrophoresis (Flu-PAGE) to detect early glycation adducts on proteins by exploiting the reversible covalent interaction between boronic acid and cis-diols that are present in fructosamine-protein adducts in glycated proteins (Pereira Morais et al., 2013). This interaction is further strengthened by the additional charge interaction between boronate and the fructosylysine amino group (Pereira Morais et al., 2010). As the anomeric cis diols produced by this interaction are absent in N- and O-glycosylation, this method enables the specific identification of glycated proteins over glycosylated and unmodified proteins as (Pereira Morais et al., 2013; Kassaar et al., 2017). This highly sensitive method detects the earliest stages of glycation, before AGEs are developed, and thus is an ideal tool for identifying reducing sugar modified proteins in complex biological samples such as plasma and brain homogenates.
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 kit Fluorescein 1 x 1mg Fluorescein-boronic acid 1 x 1mg
A strong band corresponding to glycated Human Serum Albumin is present in fluorescein-boronic acid-treated samples (HSFB) but not in control fluorescein samples (HSF). Left: Flu-PAGE analysis of normal human serum incubated with a fluorescein-boronic acid working solution made with water. Middle: Flu-PAGE analysis of normal human serum incubated with a fluorescein-boronic acid working solution made with methanol. Right panel: Coomasie Blue staining of the Flu-PAGE gel used to create the image in the centre. Flu-PAGE gels were visualized with a blue light transilluminator using an orange (595nm) filter. Normal human serum was diluted 1:10 in buffer prior to labeling with fluorescein-boronic acid or fluorescein. M – prestained molecular weight markers.
ab219802 has been referenced in 2 publications.
- Pereira Morais MP et al. Analysis of Protein Glycation Using Phenylboronate Acrylamide Gel Electrophoresis. Methods Mol Biol 1855:161-175 (2019). PubMed: 30426417
- Bartho LA et al. Maternal corticosterone in the mouse alters oxidative stress markers, antioxidant function and mitochondrial content in placentas of female fetuses. J Physiol 597:3053-3067 (2019). PubMed: 31026055