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anti-Ascorbic acid (Vitamin C) antibody

Overview

Antigen:	Ascorbic acid (Vitamin C)
Clonality:	Polyclonal
Host:	Rat
Quantity:	100 µl
Price:	624,10 $ (Plus shipping costs and VAT, Invoice in EUR)
Order number:	ABIN165502
Availability:	Ships within 5 Business days

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Additional Information » anti-Ascorbic acid (Vitamin C) antibody

Antigen	Ascorbic acid (Vitamin C)
Immunogen	Synthetic ascorbic Acid conjugated to bovine serum albumine (BSA)
Host	Rat
Clonality	Polyclonal
Format	Lyophilized
Isotype	IgG »Matching secondary antibodies
Specificity	Conjugated Ascorbic Acid (Vitamin C) . Using a conjugate ascorbic acid-protein carrier (BSA), antibody specificity was performed with an ELISA test by competition experiments with the following compounds: Compounds Cross-reactivity ratio (a) Ascorbic acid-BSA 1 Dihydroascorbic acid-BSA 1 D-isoascorbic acid-BSA 1/2,000

Application Details » anti-Ascorbic acid (Vitamin C) antibody

Application Notes	Immunohistochemistry Perfusion protocol for Adult male monkeys (Macaca fascicularis) (weight 3-3.5kg):1 - The animals can be deeply anaesthetized with ketamine (8mg/kg, intramuscular) and sodium thiopental (500mg/kg, intraperitoneal). 2 - Heparinized, and perfused via the ascending aorta with 300ml of cold physiologic saline (0.9% NaCl) and with the following fixative solutions:a) 500 ml of 1% paraformaldehyde in 0.1 M phosphate-buffer (PB), pH 7.2, at room temperature (two minutes). b) 2,500ml of 4% paraformaldehyde in 0.1 M PB, pH 7.2, at 4ºC (ten minutes). c) 5,000ml of cold 4% paraformaldehyde in 0.1 M PB, pH 7.2 (fifty minutes). d) 2,000ml of cold 5% sucrose in 0.1M PB, pH 7.2 (twenty minutes). d) Dissect out the brains and place in 10% glycerol and 2% dimethylsufoxide (DMSO) in 0.1M PB, pH 7.2, at 4ºC for two days, and finally keep at the same temperature in 20% of glycerol and 2% DMSO in PB until the brains will be cut on a freezing microtome. Around 50 µm-thick serial sections will be obtained, kept at 4ºC in PB (0.1 M, pH 7.2) containing 20% of glycerol and 30% of ethylene glycol, 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 (1h30min) and overnight at 4ºC in the same mixed solution containing anti-conjugated ascorbic acid antibodies (diluted 1/1001/5,000 as recommended dilutions). 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-rat immunogammaglobulin (Vector) diluted 1/200 in PBS. 7 - Wash during 30 min with PBS. 8 - Sections will be incubated for 1h 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).
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.
Research Area	Vitamins
Restrictions	For Research Use only