Reaction of protein amino groups with glucose leads, through the early products such as a Schiff base and Amadori rearrangement products, to the formation of advanced glycation end products (AGE). Recent immunological studies using anti-AGE antibody (6D12) demonstrated the presence of AGE-modified proteins in several human tissues: (1) human lens (nondiabetic and noncataractous), (2) renal proximal tubules in patients with diabetic nephropathy and chronic renal failure, (3) diabetic retina, (4) peripheral nerves of diabetic neuropathy, (5) atherosclerotic lesions of arterial walls, (6)β2-microglobulin forming amyloid fibrils in patients with hemodialysis-related amyloidosis, (7) senile plaques of patients with Alzheimer’s disease, (8) the peritoneum of CAPD patients, (9) skin elastin in actinic elastosis, and (10)eriod/lipofuscin deposits. These results suggest a potential role of AGE-modification in normal aging as well as age-enhanced disease processes. This antibody named as 6D12 has been used to demonstrate AGE-modified proteins in these human tissues, indicating potential usefulness of this antibody for histochemical identification and biochemical quantification of AGE-modified proteins.
AGEs Antibody Flyer [PDF]
The initial study (Ref. 1) revealed that 6D12 does not recognize early products (Schiff base and Amadori products), but shows a positive reaction to AGE-samples obtained either from proteins, lysine derivatives or monoamino-carboxylic acids, indicating the immunospecificity to a common structure among AGE-structures. The subsequent study (Ref. 10) revealed of 6D12 is an Nε- carboxymethyllysine(CML)-protein adduct.
0Immunohistochemical staining of renal proximal tubules and glomeruli in patients with diabetic nephropathy, using anti-AGE antibody 6D12
Yamada, K. et al,.Clinical nephrology, Vol.42, 354-361, 1994
Immunohistochemical staining of th eary stage of human athrosclerotic lesions of the aorta with anti-AGE antibody 6D12.
Kume, S. et al,American Journal of Pathology, Vol.147, 654-667, 1995
|Package Size||10g (40L/vial)|
|Format||Mouse monoclonal antibody 0.25 mg/mL|
|Buffer||Block Ace as a stabilizer, containing 0.1%Proclin as bacteriostat|
|Storage||Store below 20°C
Once thawed, store at 4°C. Repeated freeze-thaw cycles should be avoided.
|Purification Method||The splenic lymphocytes from BALB/c mouse, immunized with AGE-BSA were fused to myeloma P3U1 cells. The hybrid cells were screened, and the cell line (6D12) with positive reaction to AGE-human serum albumin but negative to BSA was selected through successive subclonings and grown in ascitic fluid of BALB/c mouse, from which the anti-AGE antibody was purified by Protein G affinity chromatography. (Reference No.1)|
|Application||Working dilution for immunohistochemistry: 2g /mL; for ELISA: 0.1-0.5g /mL; for WB : 0.25-5g /mL|
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2. Araki, N. et al.: Immunochemical evidence for the presence of advanced glycation end products in human lens proteins and its positive correlation with aging. J. Biol. Chem. 267: 10211, 1992.
3. Miyata, T. et al.: β2-Microglobulin modified with advanced glycation end products is a major component of hemodialysis-associated amyloidosis. J. Clin. Invest. 92: 1243, 1993.
4. Yamada, K et al.: Immunohistochemical study of human advanced glycosylation end-products (AGE) in chronic renal failure. Clin. Nephrol. 42: 354, 1994.
5. Kume, S. et al.: Immunohistochemical and ultrasturactural detection of advanced glycation end products in atherosclerotic lesions of human aorta using a novel specific monoclonal antibody. Am. J. Pathol. 147 : 654, 1995.
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9. Kimura, T. et al.: Accumulation of advanced glycation end products of the Maillard reaction with age in human hippocampal neurons. Neurosci. Lett. 208: 53,1996.
10. Ikeda, K. et al.: Nε-(carboxymethyl) lysine protein adduct is a major immunological epitope in proteins modified with advanced glycation end products of the Maillard reaction. Biochemistry 35: 8075,1996.