The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
1/500 - 1/5000.
RelevanceVasopressin, also known as arginine vasopressin (AVP) or antidiuretic hormone (ADH), is a posterior pituitary hormone that is synthesised in the hypothalamus. Vasopressin is synthesised as a precursor protein that consists of arginine vasopressin and two associated proteins, neurophysin 2 and the glycopeptide copeptin. Vasopressin, together with its carrier protein neurophysin II, is packaged into neurosecretory vesicles and transported axonally to the nerve endings in the neurohypophysis, where it is either stored or secreted into the bloodstream. Vasopressin acts as a growth factor by enhancing pH regulation through acid-base transport systems. It has a direct antidiuretic action on the kidney and also causes vasoconstriction of the peripheral vessels. Vasopressin can also contract smooth muscle during parturition and lactation. It also plays a role in cognition, tolerance, adaptation and complex sexual and maternal behaviour, as well as in the regulation of water excretion and cardiovascular functions. Mutations in the vasopressin precursor cause autosomal dominant neurohypophyseal diabetes insipidus (ADNDI), which is characterised by persistant thirst, polydipsia and polyuria.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Vasopressin antibody (ab39363)Image from Ettrup KS et al, J Chem Neuroanat. 2010 May;39(3):151-65. Epub 2009 Dec 28, Fig 2, doi:10.1016/j.jchemneu.2009.12.004.
Minipigs were deeply anesthetized with a combination of midazolam and
ketamine, prior to transcardial perfusion with phosphate buffered 4%
paraformaldehyde (pH 7.4). After perfusion, the brains were removed with special care taken to preserve the optic chiasm and the median eminence. Blocks of tissue containing the hypothalami
were dissected, postfixed in the same fixative for 1 day and subsequently cryoprotected in 30% sucrose for 3–4 days, prior to freezing. 10 series of 40-mm thick
coronal (6 animals), sagittal (1 animal), and horizontal (1 animal) cryostat
sections were collected. Coronal sections for immunohistochemistry were maintained at -18°C as free-floating sections in a cryoprotectant poly-ethylene glycol solution for up to four weeks.
Immunohistochemistry was performed using the avidin-biotin method. Accordingly,
free-floating sections were
first rinsed in Tris-buffered saline (TBS; 0.05 M; pH 7.4) for 15 minutes. Incubations with
avidin (0.1%) and biotin (0.01%) were each of 10 minute duration and each procedure
were followed by a 2 minute TBS-rinse. The sections were preincubated with 1% Triton X-
100 and 0.2% milk in TBS for 30 minutes, prior to incubation
with the primary antibody, ab39363, for 72 hours at 4°C at a 1/100 dilution. The sections were then washed three times 15 minutes
in TBS and 1% Triton X-100 prior to incubation for 1 hour at room temperature with the
secondary anti-rabbit biotinylated antibody at a 1/400 with TBS that contained 1% Triton X-
100 and 0.2% milk. After a brief rinse in TBS, the endogenous peroxidase activity was
blocked with a solution of 10 ml hydrogen peroxide, 10 ml methanol, and 80 ml TBS,
for 10 minutes. The sections were then rinsed in TBS and 1% Triton for three times 15 minutes
and incubated for 1 hour at room temperature with avidin-peroxidas diluted 1/400 in TBS that contained 1% Triton
X-100 and 0.2% milk. The sections were then rinsed three times for 15 minutes in TBS and
1% Triton, prior to avidin-peroxidase visualization. The latter step consisted of
incubation for 10 minutes in 10ml water in which one diaminobenzidine tablet had been
dissolved and to which 10 ml of 35% H2O2 had been added. Finally, the sections were
dehydrated, mounted and coverslipped with Depex. och: optic chiasm.
References for Anti-Vasopressin antibody (ab39363)
Zhao DQ & Ai HB Oxytocin and vasopressin involved in restraint water-immersion stress mediated by oxytocin receptor and vasopressin 1b receptor in rat brain. PLoS One6:e23362 (2011).
Read more (PubMed: 21858088) »