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Indications d'application
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Example of immunohistochemistry application Tissues fixation:1-Samples are fixed with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDAC) in a 4-Morpholino ethanesulfonic acid (MES 10-1M pH 6.3) buffer. 2-After dehydration with a graded series of ethanol, get the wax section. Example of immunohistochemistry 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- conjuguated Indole-3-acetic acid 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).
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Pureté
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Antiserum previously absorbed on protein carriers, and purified
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Stock
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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.
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Restrictions
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For Research Use only
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PANIER
AIDE
Antigènes
