Glutathione antibody

Details for Product No. ABIN6560000
Antigen
Reactivity
Chemical
11
5
4
3
Host
Rabbit
17
15
Clonality
Polyclonal
Conjugate
Un-conjugated
4
2
1
1
1
Application
Immunohistochemistry (IHC)
26
14
11
5
4
4
3
3
3
3
2
1
Options
Immunogen Synthetic Glutathione conjugated to bovine serum albumin (BSA)
Isotype IgG
Specificity Conjugated Glutathione . Using a conjugate Glutathione-Glutaraldehyde-Protein, antibody specificity was performed with an ELISA test by competition experiments with the following compounds: Compounds Cross-reactivity ratio (a) Glutathione-G-BSA 1 Glutathione 1/>50,000 Cysteine-G-BSA 1/>50,000 Glutamate-G-BSA 1/>50,000
Purification Antiserum previously preabsobed on protein carriers, and purified
Antigen
Target Type Chemical
Application Notes Example of Immunohistochemistry protocol Perfusion protocol 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-Heparinized, and perfused via the ascending aorta with 100 ml of cold physiologic saline (0.9% NaCl) and with the following fixative solution:a) 300 ml of cold 4% paraformaldehyde and 2% glutaraldehyde in 0.1 M phosphate-buffer (PB), pH 7.2, (two minutes). b) 600 ml of cold 4% paraformaldehyde and 2% glutaraldehyde in 0.1 M phosphate-buffer (PB), pH 7.2, (ten minutes). c) Dissect out the brains and place in a solution of 4% paraformaldehyde in 0.1M PB, pH 7.2, at 4ºC for twelve to sixteen hours. d) Before the brains will be cut on a freezing microtome, we must include the brain in growing concentrations of sucrose (a first bain of 5% of sucrose in PBS until the brains sank), after that we will repeat the same process in a solution with a higher level of sucrose (10%), 20%, 25% and finally 30%. Around 50 µm-thick serial sections will be obtained, kept at 4º C in PBS (0.1 M, pH 7.2) and processed for immunostaining. Example of immunohistochemical protocol 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 10% 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 anti-cojugated Glutathione antibodies (diluted 1/1,000 to 1/5,000 as recommended dilution). 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-rabbit 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). 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 (1/1,000-1/5,000) Recommended dilutions for Western blot (1/1,000-1/2,000)
Restrictions For Research Use only
Format Lyophilized