Malondialdehyde (MDA) is one of the major aldehyde derive from lipid peroxidation. MDA is highly reactive aldehyde and reacts with lysine residue in protein. The reaction with MDA and lysine residue leads to the formation of numerous numbers of adducts, such as dihydropyridine-lysine (DHP-lysine) type derivative. This monoclonal antibody is specific for the MDA-modified protein, especially DHP-lysine type derivative.
|Antigen:||CRA-modified keyhole-lympet hemocyanine|
|Specificity:||Specific for CRA-modified protein (especially CRA-lysine adduct)|
|Form:||Frozen (100 micro g/mL antibody in 10mM PBS containing 0.1 %NaN3 and 0.5% BSA). Purified by Protein-A.|
|Storage:||Less than -20°C|
Recommended antibody concentration is 0.5-1.0 micro gram/mL on paraformaldehyde fixed tissue.
Specifity of anti-MDA antibody.
1) Paraffin sections were deparaffinized and rehydrated. 2) Sections were quenched for several minutes with 3% hydrogen peroxide, rinsed in PBS, pretreated for 30 min with 3% nonimmune animal serum in PBS. 3) Sections were incubated overnight at 4°C with a antibody at a dilution of 1 micro gram/mL. 4) Antibody binding was visualized by the avidin-biotin-immuno peroxidase complex method using the Vectastain ABC kit (Vector, Burlingame, CA) according to the manufacturer's instructions. 5) 3,3-Diaminobenzidine tetrahydrochloride was used as the chromogen. 6) Hematoxylin or Eosin was used as the counter stain.
1) Sections from which the primary antibodies were omitted served as negative reaction controls.
2) For frozen materials, samples were fixed in 10% formalin, immersed in 30% sucrose in PBS, embedded at OCT (Sakura, Tokyo, Japan), and stored at 80°C.
3) For paraffin embedded materials, samples were fixed in 10% formalin, dehydrated, embedded in paraffin, and stored at room temperature.
4) Microwave treatment improves the target staining, whereas it declines non-specific background.
5) Protease K treatment declines the target staining
Aortic wall in the renal tissue of diabetic nephropathy or aortic wall of atherosclerosis.