Mouse Urocortin 2 ELISA Assay

$985.00

This Eagle Biosciences Mouse Urocortin 2 ELISA Assay kit is used for quantitative determination of urocortin 2 in mouse 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. Mouse urocortin 2 standard is highly purified synthetic product. This Eagle Biosciences Mouse Urocortin 2 ELISA Assay Kit is for research use only and not to be used for therapeutic procedures.

Mouse Urocortin 2 ELISA Assay

The Mouse Urocortin 2 ELISA Assay is For Research Use Only

Size: 1×96 wells
Dynamic Range: 0.82 – 100 ng/mL
Incubation Time:21 hours
Sample Type: Mouse Serum, Plasma
Sample Size: 20 µL
Alternative Name: Ucn 2


Assay 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). 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 brainstem. More recently, the expression of Ucn 2 transcripts was detected in the olfactory bulb, pituitary, cortex, hypothalamus, and spinal cord. Ucn 2 mRNA was also found to be expressed widely in a variety of peripheral tissues, most highly in the skin and skeletal muscle tissues. Ucn 2-like immunoreactivity was detected by RIA in acid extracts of mouse brain, muscle, and skin. Immunohistochemically Ucn 2 was found in both skin epidermis and adnexal structures and in the skeletal muscle myocytes. 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 pathways. 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 stress. 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.
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 treatment. 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 axis.
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 sarcopenia.
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 succeeded this time in the development of mouse urocortin 2 EIA kit which highly specific for mouse Ucn 2 with almost no crossreaction to Ucn 1 (mouse, rat), Ucn 3 (mouse), ACTH (mouse, rat) and CRF (mouse, rat, human). The kit can be used for measurement of Ucn 2 in mouse plasma or serum with high sensitivity. It will be a specifically useful tool for Ucn 2 research.


Related Products

Mouse Rat Urocortin 3 ELISA Assay Kit
Mouse Rat Urocortin 1 ELISA Assay Kit
Rat Urocortin 2 ELISA Assay Kit

Additional Information

Assay Principle


This Eagle Biosciences Mouse Urocortin 2 ELISA Assay Kit for the determination of mouse urocortin 2 in samples is based on a competitive enzyme immunoassay using combination of highly specific antibody to mouse urocortin 2 and biotin-avidin affinity system. To the wells of plate coated with rabbit anti mouse 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 o-Phenylenediamine dihydrochloride (OPD) and the concentration of mouse urocortin 2 is calculated.

Assay Procedure


  1. Before start assay, bring all the reagents and samples to room temperature (20~30℃).
  2. Add 0.35mL/well of washing solution into the wells and aspirate the washing solution in the wells. Repeat this washing procedure further twice (total 3 times). Finally, invert the plate and tap it onto an absorbent surface, such as paper toweling, to ensure blotting free of most residual washing solution.
  3. Fill 25uL of buffer solution into the wells first, then introduce 20uL of each of standard solutions (0, 0.82, 2.47, 7.41, 22.2, 66.7, 200 ng/mL) or samples and finally add 50uL of labeled antigen into the wells. The total pipetting time of standard solutions and samples for a whole plate should not exceed 30 min.
  4. Cover the plate with adhesive foil and incubate it at 4ºC overnight for 16 ~ 18 hours. (Still, plate shaker not need)
  5. After incubation, move the plate back to room temperature keeping for about 40 minutes and take off the adhesive foil, aspirate and wash the wells four times with approximately 0.35 mL/well of washing solution. Finally, invert the plate and tap it onto an absorbent surface, such as paper toweling, to ensure blotting free of most residual washing solution.
  6. Pipette 100uL of SA-HRP solution into each of the wells.
  7. Cover the plate with adhesive foil and incubate it at room temperature (20 ~ 30ºC) for 2 hours. During the incubation, the plate should be shake with a plate shaker.
    Resolve OPD tablet with 11 mL of substrate buffer. It should be prepared immediately before use.
  8. Take off the adhesive foil, aspirate and wash the wells four times with approximately 0.35 mL/well of washing solution. Finally, invert the plate and tap it onto an absorbent surface, such as paper toweling, to ensure blotting free of most residual washing solution.
  9. Add 100uL of substrate solution to each of the wells, cover the plate with adhesive foil and incubate it for 20 minutes at room temperature.
  10. Add 100uL of stopping solution into each of the wells to stop color reaction.
  11. Read the optical absorbance of the solution in the wells at 492 nm. Calculate mean absorbance values of wells containing standards and plot a standard curve on semilogarithmic graph paper (abscissa: concentration of standard; ordinate: absorbance values). Use the average absorbance of each sample to determine the corresponding value by simple interpolation from this standard curve

Typical Standard Curve


Mouse Urocortin 2 ELISA Assay

Manual

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