For Oxidative Stress Research

Anti Acrolein (ACR) monoclonal antibody


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Background

Acrolein (ACR) is a representative carcinogenic aldehyde found ubiquitously in the environment and formed endogenously through oxidation reactions, such as lipid peroxidation and myeloperoxidasecatalyzed amino acid oxidation. ACR is highly reactive aldehyde and reacts with lysine residue in protein. The reaction with ACR and lysine residue leads to the formation of numerous numbers of adducts, such as formyl-dehydropiperidino-lysine (FDP-lysine) type derivative. This antibody is specific for the ACRmodified protein, especially FDP-lysine type derivative.

Source: Mouse
Antigen: ACR-modified keyhole-lympet hemocyanine
Form: Frozen (100 micro g/mL antibody in 10mM PBS containing 0.1 %NaN3 and 0.5% BSA). Purified by Protein-A.
Application: Immunohistochemistry.
Recommended antibody concentration is 0.5-1.0 micro gram/mL on paraformaldehyde fixed tissue.
Specificity: Specific for ACR-modified protein (especially FDP-lysine type derivative)
Subclass: IgG1,kappa
Storage: Less than -20°C


Specifity of anti ACR antibody.

Immunohistochemical detection of ACR-modified protein in atherosclerotic aorta.
Koji Uchida et al.
Nagoya University

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 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.

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.

 

<Reference>
1. Protein-bound acrolein: Potential markers for oxidativestress. K.Uchida, M.Kanematsu, K.Sakai, T.Matsuda, N.Hattori,Y.Mizuno, D.Suzuki,T.Miyata, N.Noguchi, E.Niki,T.Osawa
Proc.Natl.Acad.Sci.USA,95,4882-4887(1998)
2. Protein-bound acrolein: A novel markers of oxidativestress in Alzheimer's Disease. Noel Y. Calingasan, Koji Uchida, and Gary E.Gibson
Journal of Neurochemistry.72(2),751-756(1999)
3. A 1-Hour Enzyme-Linked Immunosorbent Assay for Quantitation of Acrolein- and Hydroxynonenal- Modified Proteins by Epitope-Bound CaseinMatrix Method. K.Satoh,S.Yamada,Y.Koike,Y.Igarashi,S.Toyokuni,T.Kumano,
T.Takahata,S.Tsuchida and K.Uchida
Analytical Biochemistry.270,323-328(1999)
4. Red blood cells inhibit activation-induced cell death and oxidative stress in human peripheral blood T lymphocytes. Fonseca AM, Porto G, Uchida K, Arosa FA.
Blood. 97(10), p3152-60 (2001)
5. Thiolation of protein-bound carcinogenic aldehyde. An electrophilic acrolein-lysine adduct that covalently binds to thiols. Furuhata A, Nakamura M, Osawa T, Uchida K.
J Biol Chem. 277(31), p27919-27926 (2002)

 


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Product List

Product Name Cat# Quantity Price

Anti ACR, - (Mouse) Unlabeled DataSheet

NOF-N213310-EX

100 UG ¥144,000
$1920

Anti ACR, - (Mouse) Unlabeled DataSheet

NOF-N213320-EX

20 UG ¥36,000
$480

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