AGE (Advanced Glycation End Products) Antibody Staining Protocol for Immunohistochemistry


Description: The non enzymatic reaction of reducing carbohydrates with lysine side chains and N terminal amino groups of macromolecules (amino acids, proteins, phospholipids and nucleic acids) is called the Maillard reaction or glycation. The latter products of this process, termed advanced glycation end products (AGEs), adversely affect the functional properties of proteins, lipids and DNA. In long lived tissue proteins, these chemical modifications accumulate with age and may contribute to the pathophysiology of aging and long term complications of diabetes, atherosclerosis and renal failure.


Primary Antibody

Name: AGE Antibody

Clone: Rabbit Polyclonal

Supplier: Abcam

Catalog Number: ab23720

Dilution: 1:100 - 1:200 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 min/room temperature

Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Kidney
Fixation: Formalin-fixed paraffin sections
Positive Control: Human lens, diabetic kidney/heart/spleen/trachea/brain, arteriosclerotic plaques.
Negative Control: Omit primary antibody, isotype control, absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary



1. Kimura T et al. Accumulation of advanced glycation end products of the Maillard reaction with age in human hippocampal neurons. Neurosci Lett 208:53-6 (1996). PubMed: 8731173
2. Kume S et al. Immunohistochemical and ultrastructural detection of advanced glycation end products in atherosclerotic lesions of human aorta with a novel specific monoclonal antibody. Am J Pathol 147:654-67 (1995). PubMed: 7545874
3. Miyata T et al. beta 2-Microglobulin modified with advanced glycation end products is a major component of hemodialysis-associated amyloidosis. J Clin Invest 92:1243-52 (1993). PubMed: 8376584
4. 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 267:10211-4 (1992). PubMed: 1587810
5. Horiuchi S et al. Immunochemical approach to characterize advanced glycation end products of the Maillard reaction. Evidence for the presence of a common structure. J Biol Chem 266:7329-32 (1991). PubMed: 2019568