Collagen AGEs Assay Kit, CML-Specific, Glyoxal - Cosmo Bio Co.,Ltd.

Antibodies

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Collagen AGEs Assay Kit, CML-Specific, Glyoxal

Catalog No.: CSR-AAS-AGE-K02E
Size: 1KIT
Price: ¥80000
$1067
Storage: 4C
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component: 96-well Collagen coated plate One strip-well plate
Microplate seal 2 sheets
Anti-CML Antibodyi100Xj 100 uL
Blocking Buffer 50 mL
HRP Conjugate Secondary Antibodyi100Xj 100 ƒÊL
Sample Dilution Buffer 30 mL
Washing Bufferi10Xj 50 mL
Substrate Solution 10 mL
Stop Solution 10 mL
Glyoxal Solution 5 mL
Aminoguanidin Solutioni10 mM)
¦ positive control 250 ƒÊL
¦ The kit provides sufficient reagents to perform up to 96 assays.
Store the 96-well collagen coated plate at -20 ‹C for the long term storage.

Other:

 Background 
Although carbohydrates are indispensable for ATP production, excess amounts of carbohydrates modify amino residues of amino acids such as lysine and arginine, and results in the irreversible functional disorders of proteins by changingthe three-dimensional structure and net negative charge in patients with disordered metabolism. Since this reaction was first reported by Louis Camille Maillard in 1912, the reaction is called the Maillard reaction, or glycation. The Maillard reactionis divided by early and advanced stages. Early stage
generates Amadori rearrangement products, such as haemoglobin A1c, whereas advanced stage generates the
AGEs (advanced glycation end products), which is characterized by colour in brown andprotein
cross-linking. Collagen, the structural protein that forms skin, blood vessel wall and bone, also undergo
glycation reaction.
CML is generated by the oxidative cleavage of Amadori products by hydroxyl radicals (1), peroxynitrite
(2)and hypochloric acid (3), thus suggesting that CML is an important biological marker of oxidative stress in vivo. CML concentration, adjusted for age and duration of diabetes, is further increased in patients who have severe complications, includingnephropathy (4), retinopathy (5), and atherosclerosis (6).
CML ELISA Assay Kit provides rapid detection of CML formed by glycation with glyoxal on the collagen
coating plate. This kit is suitable to the research for functional foods and cosmeticmaterials which have anti-glycation activity.
 Principle 
Collagen coated on 96-well plate is glycated by glyoxal and CML is formed onto collagen coating wells for 24hrs at 37 ‹C. The inhibitory effects of glycation by aminoguanidin (positive control) or samples are probed with an anti-CML antibody, followed by an HRP conjugated secondary antibody. The inhibitory effect of glycation in unknown sample is determined by comparing with the inhibitory effect of glycation by aminoguanidin (positive control).
gCollagen AGEs Assay Kit, CML-Specific, Glyoxalh is suitable to the research for functional foods and cosmetic materials which have an anti-glycation activity.



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Production pathway of AGEs generating aldehyde

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Inhibitory effect of aminoguanidine and phytochemicals on the CML formation. (10 mM Glyoxal, glycation for 1 day)

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Reference:

•  Nagai R. et al., Biochem. Biophys. Res. Commun. 234, 167-172 (1997)
•  Nagai R. et al., Diabetes. 51, 2833-2839 (2002)
•  Mera K. et al., Free Radic. Res. 41, 713-718 (2007)
•  Mera K. et al., J. Immunol. Methods 334(1-2), 82-90 (2008)
 
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