Vasopressin Antibody (Rabbit mAb) [P14E10]

Application: Reactivity: Mouse, Rat, Human

Usage Information

Dilution
WB1:1000 IP1:30 IHC1:2000 FCM1:500

Application
WB, IP, IHC, FCM

Reactivity
Mouse, Rat, Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
17 kDa

Positive Control	Mouse brain tissue; Rat brain tissue; Rat hypothalamus tissue; Mouse hypothalamus tissue; HEK-293T cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
Vasopressin Antibody (Rabbit mAb) [P14E10] detects Vasopressin.

Clone
P14E10

Synonym(s)
Vasopressin-neurophysin 2-copeptin, AVP-NPII, Avp

Background
Arginine vasopressin, also known as antidiuretic hormone, is a cyclic nonapeptide synthesized as part of a larger precursor alongside its carrier protein neurophysin II, which binds vasopressin during axonal transport and storage within the neurohypophysis before hormone release into the circulation. Vasopressin signals through three related G-protein-coupled receptor subtypes, V1aR, V1bR, and V2R, and additionally cross-activates the structurally related oxytocin receptor, with receptor subtype determining which downstream second-messenger cascade is engaged. The V2 receptor is unique among the vasopressin receptor subtypes in coupling to the stimulatory G protein Gs, and vasopressin binding to V2R on the basolateral membrane of kidney collecting duct principal cells activates adenylyl cyclase, raising intracellular cyclic AMP and activating protein kinase A; PKA is tethered specifically to aquaporin-2-bearing intracellular vesicles through A-kinase anchoring proteins, including AKAP18delta and AKAP220, and this anchored PKA population phosphorylates the water channel aquaporin-2, most notably at serine 256, driving redistribution of aquaporin-2 vesicles from the cytoplasm to the apical plasma membrane. This translocation of aquaporin-2 completes the water permeability pathway, since basolateral aquaporin-3 and aquaporin-4 are constitutively expressed and provide the exit route for water that enters through apical aquaporin-2, together enabling net water reabsorption and urine concentration. Phosphoproteomic analysis in PKA-null collecting duct cells shows that V2R-mediated vasopressin signaling is predominantly but not entirely PKA-dependent, with residual, PKA-independent phosphorylation events detected at aquaporin-2 serine 256 and in downstream kinases including SPAK and an atypical protein kinase C, indicating that a secondary, PKA-independent signaling arm contributes to the trafficking response. Beyond the cAMP-PKA axis, V2R activation also engages arrestin-dependent pathways that drive receptor internalization and MAP kinase phosphorylation linked to cell growth and proliferation, distinguishing acute water-channel trafficking from longer-term proliferative signaling downstream of the same receptor. In contrast to V2R, the V1a and V1b receptor subtypes couple to Gq and activate phospholipase C, generating inositol trisphosphate and diacylglycerol that mobilize intracellular calcium and activate protein kinase C, a signaling route responsible for vasopressin's vasoconstrictive action on peripheral vessels rather than its renal antidiuretic effect.

Background

Arginine vasopressin, also known as antidiuretic hormone, is a cyclic nonapeptide synthesized as part of a larger precursor alongside its carrier protein neurophysin II, which binds vasopressin during axonal transport and storage within the neurohypophysis before hormone release into the circulation. Vasopressin signals through three related G-protein-coupled receptor subtypes, V1aR, V1bR, and V2R, and additionally cross-activates the structurally related oxytocin receptor, with receptor subtype determining which downstream second-messenger cascade is engaged. The V2 receptor is unique among the vasopressin receptor subtypes in coupling to the stimulatory G protein Gs, and vasopressin binding to V2R on the basolateral membrane of kidney collecting duct principal cells activates adenylyl cyclase, raising intracellular cyclic AMP and activating protein kinase A; PKA is tethered specifically to aquaporin-2-bearing intracellular vesicles through A-kinase anchoring proteins, including AKAP18delta and AKAP220, and this anchored PKA population phosphorylates the water channel aquaporin-2, most notably at serine 256, driving redistribution of aquaporin-2 vesicles from the cytoplasm to the apical plasma membrane. This translocation of aquaporin-2 completes the water permeability pathway, since basolateral aquaporin-3 and aquaporin-4 are constitutively expressed and provide the exit route for water that enters through apical aquaporin-2, together enabling net water reabsorption and urine concentration. Phosphoproteomic analysis in PKA-null collecting duct cells shows that V2R-mediated vasopressin signaling is predominantly but not entirely PKA-dependent, with residual, PKA-independent phosphorylation events detected at aquaporin-2 serine 256 and in downstream kinases including SPAK and an atypical protein kinase C, indicating that a secondary, PKA-independent signaling arm contributes to the trafficking response. Beyond the cAMP-PKA axis, V2R activation also engages arrestin-dependent pathways that drive receptor internalization and MAP kinase phosphorylation linked to cell growth and proliferation, distinguishing acute water-channel trafficking from longer-term proliferative signaling downstream of the same receptor. In contrast to V2R, the V1a and V1b receptor subtypes couple to Gq and activate phospholipase C, generating inositol trisphosphate and diacylglycerol that mobilize intracellular calcium and activate protein kinase C, a signaling route responsible for vasopressin's vasoconstrictive action on peripheral vessels rather than its renal antidiuretic effect.

References
https://pubmed.ncbi.nlm.nih.gov/32219907/ https://pubmed.ncbi.nlm.nih.gov/19096775/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%