Anti Crotonaldehyde antibody
(Anti CRA monoclonal antibody)
FOR RESEARCH USE ONLY
About crotonaldehyde (CRA)
Crotonaldehyde (CRA) is a representative carcinogenic aldehyde formed endogenously through lipid
peroxidation. CRA is highly reactive aldehyde and reacts with lysine residue in protein. The reaction with CRA
and lysine residue leads to the formation of numerous numbers of adducts. This antibody is specific for the
CRA-modified protein.
Specifications
Subclass:
Mouse IgG2b (lambda), clone 82D3
Antigen:
CRA-modified keyhole-lympet hemocyanine
Specificity:
Specific for CRA-modified protein (especially CRA-lysine adduct)
Form:
Frozen (100 µg/mL antibody in 10mM PBS containing 0.1% NaN3 and 0.5% BSA).
Purified by Protein-A.
Storage:
Less than -20°C
Application:
Immunohistochemistry.
Recommended antibody concentration is 0.5-1.0 µg/mL on paraformaldehyde fixed tissue.
Reference
1)
Endogenous formation of protein adducts with carcinogenic aldehydes.
K Ichihashi, T Osawa, S Toyokuni, K Uchida J Biol Chem 276(26), p23903-23913 (2001)
Product name
Code
Content
Anti Crotonaldehyde (CRA) monoclonal antibody
MCA-030n
30 µg IgG
Manufactured by NOF corporation, Japan.
FOR RESEARCH USE ONLY. Not for diagnostic, medical or other use.
Immunohistochemistry
Procedure
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 µg/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.
Technical Note
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.
Positive control
Aortic wall in the renal tissue of diabetic nephropathy or
aortic wall of atherosclerosis.
FOR RESEARCH USE ONLY. Not for diagnostic, medical or other use.
