Technical datasheet
Description
The monoclonal antibody CML26 recognizes human CML (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.
Cross Reactivity
| Cross reactant |
Reactivity |
| Multispecies |
Yes |
Immunogen
CML-KLH
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin and 0.02% sodium azide
Species
Mouse IgG1 , predominantly. Other isotypes maybe present.
Application
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F
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FC
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FS
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IA
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IF1
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IP
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P1-8
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W2,7
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Yes
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No
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N.D.
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N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot
Application IA has been tested by Hycult Biotech. Application F is based on personal communication.
Application notes
P: fixation in 4% formalin; cardiac tissue sections (4 mm) deparaffinised for 10 min in xylene at room temperature, dehydrated by decreasing ethanol. Sections stained with haematoxylin and eosin. Blocking endogenous peroxidase activity with 0.3% hydrogen peroxide in methanol for 30 min. No heating to prevent artificial induction of CML. (Ref 1)
IF: 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. 0.5–3.0- mm-thick sections were cut with a glass knife.(Ref 1)
Use
For immunohistochemistry, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50. For functional studies, in vitro dilutions have to be optimized in user’s experimental setting.
Positive control
Intramyocardial arteries
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least one year. The exact expiry date is indicated on the label.
References
- Bruynzeel, AM. Anti-inflammatory agents and monoHER protect against DOX-induced cardiotoxicity and accumulation of CML in mice. B J of Cancer 2007: 96, 937
- Ciapaite J. Functioning of oxidative phosphorylation in liver mitochondria of high-fat diet fed rats. Biochim Biophys Acta. 2007, 1772:307.
- Baidoshvili A. N(varepsilon)-(carboxymethyl)lysine depositions in intramyocardial blood vessels in human and rat acute myocardial infarction: a predictor or reflection of infarction?Arterioscler Thromb Vasc Biol. 2006, 26:2497.
- Lieuw-a-Fa ML. Interaction of Nepsilon(carboxymethyl)lysine- and methylglyoxal-modified albumin with endothelial cells and macrophages. Splice variants of RAGE may limit the responsiveness of human endothelial cells to AGEs. Thromb Haemost. 2006, 95:320.
- van Heijst JW. Argpyrimidine-modified Heat shock protein 27 in human non-small cell lung cancer: a possible mechanism for evasion of apoptosis.Cancer Lett. 2006, 241:309.
- Sommeijer DW. More fibrosis and thrombotic omplications but similar expression patterns of markers for coagulation and inflammation in symptomatic plaques from DM2 patients. J Histochem Cytochem. 2004 , 52:1141
- Schalkwijk CG. Increased accumulation of the glycoxidation product Nepsilon-(carboxymethyl) lysine in hearts of diabetic patients: generation and characterisation of a monoclonal anti-CML antibody. Biochim Biophys Acta. 2004, 221636:2
- Baidoshvili A. N(omega)-(carboxymethyl)lysine depositions in human aortic heart valves: similarities with atherosclerotic blood vessels. Atherosclerosis. 2004, 174:287.
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.
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