What is Methylglyoxal ?
Methylglyoxal (MG), an endogenous metabolite that increases in diabetes and is a common intermediate in the Maillard reaction (glycation), reacts with proteins and forms advanced glycation end products (AGE). MG reacts with arginine residue in protein and forms numerous numbers of adducts, such as argpyrimidine. This antibody is specific for argpyrimidine: N-(5-hydroxy-4,6-dimethylpyrimidine-2-yl)-L-ornithine.
Specifity of anti MG antibody.
Immunohistochemical detection of MG adducts in atherosclerotic aorta.
Noriyuki Shibata et al.
Tokyo Women’s medical University
1) Sections from which the primary antibodies were omitted served as negative reaction controls.
2) For frozen materials, samples were fixed in 10% formalin, immersed in 30% sucrose in PBS, embedded at OCT (Sakura, Tokyo, Japan), and stored at 80°C.
3) For paraffin embedded materials, samples were fixed in 10% formalin, dehydrated, embedded in paraffin, and stored at room temperature.
4) Microwave treatment improves the target staining, whereas it declines non-specific background. 5) Protease K treatment declines the target staining.
1) Paraffin sections were deparaffinized and rehydrated.
2) Sections were quenched for several minutes with 3% hydrogen peroxide, rinsed in PBS, pretreated for 30 min with 3% nonimmune animal serum in PBS.
3) Sections were incubated overnight at 4°C with a antibody at a dilution of 1 micro gram/mL.
4) Antibody binding was visualized by the avidin-biotin-immuno peroxidase complex method using the Vectastain ABC kit (Vector, Burlingame, CA) according to the manufacturer's instructions.
5) 3,3-Diaminobenzidine tetrahydrochloride was used as the chromogen.
6) Hematoxylin or Eosin was used as the counter stain.
Aortic wall in the renal tissue of diabetic nephropathy or aortic wall of atherosclerosis