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useful for measurement of Urocortin 2 in rat plasma or serum with high sensitivity

Rat Urocortin 2 EIA Kit

Background

Urocortin 2 (Ucn 2), also known as stresscopin-related peptide, is a novel predicted neuropeptide related to corticotropin-releasing factor (CRF). The peptide consisting of 38 amino acid residues was first demonstrated to be expressed centrally and to bind selectively to type 2 CRF receptor (CRFR2)1). In the rodent, Ucn 2 transcripts were shown to be expressed in the discrete regions of the central nervous system including stress-related cell groups in the hypothalamus and brainstem1). More recently, the expression of Ucn 2 transcripts was detected in the olfactory bulb, pituitary, cortex, hypothalamus, and spinal cord2). Ucn 2 mRNA was also found to be expressed widely in a variety of peripheral tissues, most highly in the skin and skeletal muscle tissues3). Ucn 2-like immunoreactivity was detected by RIA in acid extracts of mouse brain, muscle, and skin3). Immunohistochemically Ucn 2 was found in both skin epidermis and adnexal structures and in the skeletal muscle myocytes3). Ucn 2 gene transcription was stimulated in the hypothalamus and brainstem by glucocorticoid administration to the mouse and inhibited by removal of glucocorticoids by adrenalectomy, suggesting a putative link between the CRFR1 and CRFR2 pathways2). On the other hand, in the rat a stressor-specific regulation of Ucn 2 mRNA expression in the hypothalamic paraventricular nucleus was demonstrated, which raised the possibility of a modulary role of Ucn 2 mRNA in stress-induced alteration of anterior and posterior pituitary function, depending on the type of stress4). Administration of dexamethasone to the mouse resulted in a decrease of Ucn 2 mRNA levels in the back skin region. Adrenalectomy signifcantly increased Ucn 2 mRNA levels in the skin, and the levels were reduced back to normal levels after corticoid replacement 3).

CRFR2 is found in cardiomyocytes and in endothelial and smooth muscle cells of the systemic vasculature. Ucn 2 is expressed in the mouse cardiomyocytes. In the mouse, Ucn 2 treatment augmented heart rate, exhibited potent inotropic and lusitropic actions on the left ventricle, and induced a downward shift of the diastolic pressure-volume relation5). Ucn 2 also reduced systemic arterial pressure, associated with a lowering of systemic arterial elastance and systemic vascular resistance. The effects of Ucn 2 were specific to CRFR2 function and independent of beta-adrenergic receptors. These experiments demonstrated the potent cardiovascular physiologic actions of Ucn 2 in the both wild-type and cardiomyopathic mice and support a potential beneficial use of Ucn 2 in congestive heart failure treatment5). The use of Ucn 2 was also proposed to treat ischemic heart disease because of its potent cardioprotective effect in the mouse heart and its minimal impact on the hypothalamic stress axis6).

Administration of Ucn 2 to the mouse prevented the loss of skeletal muscle mass resulting from disuse due to casting, corticosteroid treatment, and nerve damage. In addition, Ucn 2 treatment prevented the loss of skeletal muscle force and myocyte cross-sectional area that accompanied muscle mass losses resulting from disuse due to casting. In normal muscles of the mouse, Ucn 2 increased skeletal muscle mass and force. It was thus proposed that Ucn 2 might find utility in the treatment of skeletal muscle wasting diseases including age-related muscle loss or sarcopenia7).

Mouse urocortin 2 (Ucn 2) is a new peptide predicted from mouse cDNA sequence and its physiologic and pathophysiologic significance has not yet been fully elucidated. However, the experimental data presented to date provided evidence for the important physiologic roles of Ucn 2 and urge the necessity of further investigation of the peptide from various points of view.

We have already developed mouse urocortin 2 (Ucn2) EIA kit (YK190) and mouse urocortin 3 (Ucn3) EIA (YK200). This time, as a part of tools for urocortin research, our laboratory developed rat urocortin 2 (Ucn2) EIA kit (YK191), which highly specific for rat Ucn 2 with almost no crossreaction to Ucn 1 (mouse, rat), Ucn 2 (mouse), Ucn 3 (mouse), and CRF (mouse, rat, human). The kit can be used for measurement of Ucn 2 in rat plasma or serum with high sensitivity. It will be a specifically useful tool for Ucn 2 research.

<Reference> ReferencePDF

Features

This EIA kit is used for quantitative determination of urocortin 2 in rat plasma & serum samples. The kit is characterized by its sensitive quantification and high specificity. In addition, it has no influence by other components in samples. Rat urocortin 2 standard is highly purified synthetic product.

・The assay kit can measure rat urocortin 2 within the range of 1.56-100 ng/mL.
・The assay is completed within 16-18 hr.+3 hr
・With one assay kit, 40 samples can be measured in duplicate.

・Test sample : Rat plasma & serum Sample volume  50 μL

・The 96-wells plate in kit was consisted by 8-wells strips, and the strips can be used separately.

・Precision and reproducibility
Intra-assay CV(%)   Rat plasma 4.04-11.09   Rat serum 5.19-10.59
Inter-assay CV(%)   Rat plasma 2.35-11.44   Rat serum 4.72-7.88

・Stability and Storage
Store all of the components at 2-8°C.
6 months from the date of manufacturing. The expiry date is described on the label of kit.

<Specificity>
The EIA kit has high specificity to rat urocortin 2 and shows no crossreactivity to urocortin 1 (mouse, rat), urocortin 2 (mouse), urocortin 3 (mouse), nor CRF (mouse, rat, human).

<Test Principle>
This EIA kit for determination of rat urocortin 2 in samples is based on a competitive enzyme immunoassay using combination of highly specific antibody to rat urocortin 2 and biotin-avidin affinity system. To the wells of plate coated with rabbit anti rat urocortin 2 antibody, standard or samples, labeled antigen are added for competitive immunoreaction. After incubation and plate washing, horse radish peroxidase (HRP) labeled streptoavidin (SA) is added to form HRP labeled streptoavidin-biotinylated antigen-antibody complex on the surface of the wells. Finally, HRP enzyme activity is determined by 3,3’,5,5’-Tetramethylbenzidine (TMB) and the concentration of rat urocortin 2 is calculated

Composition

  Form Quantity Main Ingredient
1.Antibody coated plate Microtiter plate 1 plate
(96 wells)
Rabbit anti mouse / rat urocortin 2 antibody
2.Standard antigen lyophilized 1 vial
(100ng)
Synthetic rat urocortin 2 (100ng/vial)
3.Labeled antigen lyophilized 1 vial Biotinylated rat urocortin 2
4.SA-HRP solution Liquid 1 bottle
(12 mL)
HRP labeled streptavidin
5.Enzyme substrate solution Liquid 1 bottle
(12 mL)
3, 3’,5 ,5’-Tetramethylbenzidine (TMB)
6.Stopping solution Liquid 1 bottle
(12 mL)
1 M H2SO4
7.Buffer solution Liquid 1 bottle
(25 mL)
Citrate buffer
8.Washing solution (Concentrated) Liquid 1 bottle
(50 mL)
Concentrated saline
9.Adhesive plate seal   3 pieces  

Product List

Product Name Cat# Quantity Price

Urocortin 2 EIA kit

YII-YK191-EX 1KIT

¥ 76,000
$ 1014
€ 760

Chromogranin A EIA

YII-YK070-EX 1KIT

¥ 76,000
$ 1014
€ 760

Urocortin 2, EIA

YII-YK190-EX 1KIT

¥ 76,000
$ 1014
€ 760

Urocortin 3, EIA Kit

YII-YK200-EX 1KIT

¥ 76,000
$ 1014
€ 760

Anti Urocortin

YII-Y361-EX 50UL

¥ 35,000
$ 467
€ 350

Anti Urocortin 1

YII-Y362-EX 50UL

¥ 35,000
$ 467
€ 350

Anti Urocortin 2

YII-Y363-EX 50UL

¥ 35,000
$ 467
€ 350

Anti Urocortin 3

YII-Y364-EX 50UL

¥ 35,000
$ 467
€ 350