Acrolein antibody

Antigen: Acrolein

Clonality: Polyclonal

Host: Rabbit

Application: Immunohistochemistry (IHC)

Catalog no.: ABIN94583

Additional Information

Immunogen: Synthetic Acrolein conjugated to bovine serum albumin (BSA)

Format: Lyophilized

Isotype: IgG  (Matching secondary antibodies)

Specificity: Conjugated Acrolein . Using a conjugate Acrolein-Protein, antibody specificity was performed with an ELISA test by competition experiments with the following compounds: Compounds Cross-reactivity ratio (a) Acrolein-BSA 1 Malondialdehyde-BSA 1/55 Glyoxal-BSA 1/>50,000 Formaldehyde-BSA 1/>50,000 Glutaraldehyde-BSA 1/>50,000 (a): Acrolein-BSA concentration /Conjugated closed related compounds concentration at half displacement. BSA = Bovine Serum Albumin.

Application Details

Application Notes: Example of immunohistochemistry protocol Perfusion ( Example for Adult male Sprague Dawley (weight around 0.5 kg)). 1-The animals can be deeply anaesthetized (for example with urethane-0.5-1.5g/kg, intraperitoneal). 2-Perfused via the ascending aorta with 50 ml of NaCl 9g/l (Heparinized) and pass trhough the system 800-1000 ml of cold 4% paraformaldehyde (Merck) in 0.1 M PB, pH 7.2-7.4, (ten minutes). 3-Dissect out the organs and place in a solution of 4% paraformaldehyde in 0.1M PB, pH 7.2, at 4ºC for twelve to sixteen hours. Immunohistochemistry 1-In order to avoid possible interference with endogenous peroxidase, free-floating sections will be treated with distilled water containing NH 3 (20%), H 2 O 2 (30%) and NaOH (1%) for 20 min (other method is using a solution with 33% of H 2 O 2 and 66% of methanol). 2-Then, wash the sections for 20 min in 0.15 M phosphate-buffered saline (PBS) (pH 7.2) 3-Pre-incubate for 30 min in PBS containing 2-10% (variable to adjust) of normal horse serum and 0.3% of Triton X-100 (mixed solution). 4-Incubate at room temperature (1h 30min) and overnight at 4º C in the same mixed solution containing the diluted (1/500-1/1000) antiserum (AP051). 5-Then, the sections will be wash in PBS (30 min). 6-After that we will incubate for 60 min at room temperature with biotinylated anti-(species) immunogammaglobulin (Vector) diluted 1/200 in PBS. 7-Wash during 30 min with PBS. 8-Sections will be incubated for 1 h with a 1/100 diluted avidin-biotin-peroxidase complex (Vectastain) in the mixed solution. 9-After that we will wash the sections in PBS (30 min) 10-Wash with Tris-HCl buffer (pH 7.6)(10 min). 11-The tissue-bound peroxidase will be developed with H 2 O 2 using 3, 3' diaminobenzidine as chromogen. 12-Finally the sections will be rinsed with PBS and coverslipped with PBS/Glycerol (1/1). Recommended dilutions for Immunohistochemistry (1/500-1/1,000) Recommended dilutions for Western blot (1/1,000 ­ 1/2,000)

Purity: Antiserum previously preabsobed on protein carriers, and purified

Storage: After reconstitution with 50µl of distilled water and 50µl of glycerol, the aliquot can be repeated freezed (up to five times), and stable at least 2 years

Restrictions: For Research Use only

Publications

Publications: Shen, Yang, Dong et al.: "Oxidative damage is a potential cause of cone cell death in retinitis pigmentosa." in: Journal of cellular physiology, Vol. 203, Issue 3, pp. 457-64, 2005 (PubMed).

Komeima, Rogers, Lu et al.: "Antioxidants reduce cone cell death in a model of retinitis pigmentosa." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 103, Issue 30, pp. 11300-5, 2006 (PubMed).