CML (Carboxymethyl-lysine, Ne-carboxymethyl Lysine)

CML is known to be formed from the oxidation of both carbohydrates and lipids. This makes CML a biomarker of general oxidative stress. Carboxymethyl-lysine (CML) is a well-characterized glycoxidation product that accumulates in tissues with age, and its rate of accumulation is accelerated in diabetes. Glycoxidation products are a subset of advanced glycation endproducts (AGEs) that are formed by the nonenzymatic glycation and subsequent irreversible oxidation of proteins. Oxidative stress and protein modification have been implicated in the pathogenesis of the chronic complications of diabetes, including nephropathy and atherosclerosis. The accumulation of CML in long-lived tissue such as skin collagen reflects oxidative stress over an extended period of the life-span, and has been shown to be greater in patients with diabetic complications than those without complications. Applications: Suitable for use in Western Blot, ELISA, Immunofluorescence and Immunohistochemistry. Other applications not tested. Recommended Dilution: Immunohistochemistry (Paraffin): fixation in 4% formalin; cardiac tissue sections (4mm) deparaffinised for 10min in xylene at room temperature, dehydrated by decreasing ethanol. Sections stained with haematoxylin and eosin. Blocking endogenous peroxidase used Immunofluorescence: After fixation in 2% phosphate-buffered glutaraldehyde solution the heart tissue was post-fixed in 1% osmium tetroxide. The tissue was dehydrated through a graded series of ethanol. 0.5-3-mm-thick sections were cut with a glass knife. Optimal dilutions to be determined by the researcher. Positive Control: Intramyocardial arteries Storage and Stability: May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.