Carboxymethyl Lysine Antibody (Mouse mAb) [B15G14]

Application: Reactivity: All

Usage Information

Dilution
WB1:1000 IHC1:50 FCM1:2000

Application
WB, IHC, FCM, ELISA

Reactivity
All

Source
Mouse Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Positive Control	HepG2 cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
Carboxymethyl Lysine Antibody (Mouse mAb) [B15G14] detects endogenous levels of total Carboxymethyl Lysine modified protein.

Clone
B15G14

Synonym(s)
CML, N Epsilon (Carboxymethyl) Lysine

Background
Carboxymethyl lysine, also designated Nε-carboxymethyllysine or CML, is a non-enzymatic modification of the lysine side chain and represents the most extensively characterized advanced glycation end product, formed when reducing sugars react with free amino groups on proteins through the Maillard reaction. CML arises through three convergent chemical routes: oxidative cleavage of the Amadori rearrangement product formed early in the glycation reaction, fragmentation of the Schiff base intermediate through the Namiki pathway, and direct reaction of lysine residues with glyoxal generated during glucose autoxidation, with modeling of these competing pathways indicating that the glyoxal route dominates CML generation under most physiological glucose and oxidative conditions. Once formed, CML functions as a ligand for the receptor for advanced glycation end products, RAGE, a pattern-recognition receptor expressed on adipocytes, endothelial cells, and macrophages, and CML engagement of RAGE activates downstream signaling that converges on nuclear factor-κB, driving production of inflammatory mediators. In adipose tissue, RAGE-mediated accumulation of CML activates this CML-RAGE axis to dysregulate adipokine expression, and this mechanism links elevated CML directly to obesity-associated insulin resistance, positioning the CML-RAGE interaction as a specific molecular route connecting a glycation product to a defined metabolic disease phenotype rather than a generalized marker of tissue damage. Beyond adipose tissue, CML-RAGE engagement activates the ERK and NF-κB pathways to drive migration, invasion, and stemness in osteosarcoma cells, and CML accumulation correlates with advanced tumor stage, indicating that the same receptor-coupled signaling cascade operates across metabolic and oncological contexts depending on the tissue expressing RAGE. CML also accumulates in tissues under conditions of oxidative stress and elevated glucose, including diabetic hearts and fatty liver, where its endogenous formation induces inflammatory marker expression independent of dietary intake, while dietary CML formed during thermal food processing contributes an additional, exogenous source that adds to the endogenous pool. Because CML formation depends on reactive carbonyl intermediates including glyoxal, methylglyoxal, and glucosone rather than on a single fixed precursor, its generation is sensitive to oxidative conditions, transition metal availability, and thermal processing, making CML both a chemically well-defined marker of cumulative glycoxidative burden and a functionally active RAGE ligand relevant to researchers studying diabetic complications, obesity-associated inflammation, and RAGE-driven tumor signaling.

Background

Carboxymethyl lysine, also designated Nε-carboxymethyllysine or CML, is a non-enzymatic modification of the lysine side chain and represents the most extensively characterized advanced glycation end product, formed when reducing sugars react with free amino groups on proteins through the Maillard reaction. CML arises through three convergent chemical routes: oxidative cleavage of the Amadori rearrangement product formed early in the glycation reaction, fragmentation of the Schiff base intermediate through the Namiki pathway, and direct reaction of lysine residues with glyoxal generated during glucose autoxidation, with modeling of these competing pathways indicating that the glyoxal route dominates CML generation under most physiological glucose and oxidative conditions. Once formed, CML functions as a ligand for the receptor for advanced glycation end products, RAGE, a pattern-recognition receptor expressed on adipocytes, endothelial cells, and macrophages, and CML engagement of RAGE activates downstream signaling that converges on nuclear factor-κB, driving production of inflammatory mediators. In adipose tissue, RAGE-mediated accumulation of CML activates this CML-RAGE axis to dysregulate adipokine expression, and this mechanism links elevated CML directly to obesity-associated insulin resistance, positioning the CML-RAGE interaction as a specific molecular route connecting a glycation product to a defined metabolic disease phenotype rather than a generalized marker of tissue damage. Beyond adipose tissue, CML-RAGE engagement activates the ERK and NF-κB pathways to drive migration, invasion, and stemness in osteosarcoma cells, and CML accumulation correlates with advanced tumor stage, indicating that the same receptor-coupled signaling cascade operates across metabolic and oncological contexts depending on the tissue expressing RAGE. CML also accumulates in tissues under conditions of oxidative stress and elevated glucose, including diabetic hearts and fatty liver, where its endogenous formation induces inflammatory marker expression independent of dietary intake, while dietary CML formed during thermal food processing contributes an additional, exogenous source that adds to the endogenous pool. Because CML formation depends on reactive carbonyl intermediates including glyoxal, methylglyoxal, and glucosone rather than on a single fixed precursor, its generation is sensitive to oxidative conditions, transition metal availability, and thermal processing, making CML both a chemically well-defined marker of cumulative glycoxidative burden and a functionally active RAGE ligand relevant to researchers studying diabetic complications, obesity-associated inflammation, and RAGE-driven tumor signaling.

References
https://pubmed.ncbi.nlm.nih.gov/24723555/ https://pubmed.ncbi.nlm.nih.gov/26462729/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%