Urocortin (Urocortin1: Ucn1) is first identified in rat 1), and later in human2) and mouse3). It is the second mammalian member of the CRF family. Rat and mouse Ucn1 have the same amino acid sequence and display 95% structure homology to human Ucn1, 45% to CRF and 63% to urotensin.
In the rat, Ucn1 immunoreactivity (IR) was shown to distribute widely in central nervous system, endocrine organs, and digestive system and its concentration was highest in pituitary (11 pmol/g, w.w.) 4). Koziez, Yanaihara et al. used a polyclonal antibody against rat Ucn1 to define the distribution of Ucn1-IR in rat central nervous system and found a large number of neurons with Ucn1-IR in rat brain 5).
Synthetic human Ucn1 binds with high affinity to CRF receptor type 1(CRFR1), 2 alpha(CRFR2α) and 2 beta (CRFR2β). CRFR1 and CRFR2 have been shown to link to the development of stress-related disorders, such as mood and eating disorders. CRFR1 is expressed predominantly in the brain and pituitary, whereas CRFR2 expression is limited to particular brain areas and to some peripheral organs 6). Data were also presented supporting the hypothesis that this peptide is the endogenous ligand for the CRFR2. 5)
Synthetic human Ucn1 stimulates cAMP accumulation in cells stably transfected with those receptors and acts in vitro to release ACTH from dispersed rat anterior pituitary cells. In addition, the CRF-binding protein binds human Ucn1 with high affinity and can prevent Ucn1-stimulated ACTH secretion in vitro2). Ucn1 was suggested to play important roles in various tissues in normal rats, but shown not to behave as a hypothalamic hypophysiotropic factor in mediating adrenocorticotropin secretin in adrenalectomized rats 4). Ucn1 has been implicated in various endocrine responses, such as blood pressure regulation, as well as in higher cognitive functions 5).
Synthetic human Ucn1 also stimulates plasma ACTH, cortisol and atrial natriuretic peptide (ANP) secretin and suppresses plasma ghrelin in healthy male volunteers7). In the human, plasma Ucn1 is elevated in heart failure, especially in its early stages. This fact may useful in the diagnosis of early heart failure 8).
We have already developed mouse urocortin 2 (Ucn2) EIA kit (YK190), rat urocortin 2 (Ucn2) EIA kit (YK191) and mouse/rat urocortin 3 (Ucn3) EIA kit (YK200). This time, as a part of tools for urocortin research, our laboratory developed mouse/rat Ucn1 EIA kit (YK210), which is highly specific for mouse/rat Ucn1 with almost no crossreaction with Ucn2 (mouse), Ucn2 (rat), Ucn3 (mouse, rat), ACTH (mouse, rat), ACTH (human) and CRF (mouse, rat, human). The kit can be used for measurement of Ucn1 in mouse/rat plasma or serum with high sensitivity. It will be a specifically useful tool for Ucn1 research.
1. Vzquez-Palacios G, Retana-Mrquez S et al: Further definition of the effect of corticosterone on the sleep-wake pattern in the male rat. Pharmacol Biochem Behav 70:305-310, 2001
2. Goymann W, Mstl E and Gwinner E: Corticosterone metabolites can be measured noninvasively in excreta of European Stonechats (Saxicola torquata rubicola). The Auk 119:1167-1173. 2002
3. Hupe JM, James AC et al: Cortical feedback improves discrimination between figure and background by V1, V2 and V3 neurons.Nature 394:784-787, 1998
4. Kitaysky AS, Kitaiskaiya EV et al: Dietary restriction causes chronic elevation of corticosterone and enhances stress response in red-legged kittiwake chicks. J Comp Physiol B: Regulatory, Integrative and Comparative Physiology 171:701-709, 2001
5. Thelling O, Noel G et al: Stress hormone secretion and gut signal transducer (STAT) proteins after burn injury in rats. Shock 16:393-397, 2001
6. National Committee for Clinical Laboratory Standards Evaluation Protocols, SC1 (1989), Vallanova, PA: NCCLS
This EIA kit is used for quantitative determination of Ucn 1 in mouse/rat plasma and 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/rat Ucn1 standard is highly purified synthetic product.
・The assay kit range: 1.563-100 ng/m
・Test sample: mouse/rat plasma or serum
・The 96-wells plate of this kit consists of 12 8-wells strips, so that divided use by the strips is possible at your option.
・Precision and reproducibility
・Stability and Storage
The EIA kit has high specificity to mouse/rat Ucn1 and shows crossreactivity with none of Ucn2 (mouse), Ucn2 (rat), Ucn3 (mouse, rat), ACTH (mouse, rat), ACTH (human) and CRF (mouse, rat, human).
This EIA kit for determination of mouse/rat Ucn1 in samples is based on a competitive enzyme immunoassay using combination of highly specific antibody to mouse/rat Ucn1 with biotin-avidin affinity system. The 96-wells plate is coated with rabbit anti-mouse/rat Ucn1 antibody. Mouse/rat Ucn1 standard or samples and biotin labeled antigen are added to the wells for competitive immunoreaction. After incubation and plate washing, horse radish peroxidase (HRP) labeled streptoavidin (SA) is added to form HRP labeled SA-biotinylated mouse/rat Ucn1-antibody complex on the surface of the wells. Finally, HRP enzyme activity is determined by 3,3’,5,5’-Tetramethyl benzidine (TMB) and the concentration of mouse/rat Ucn1 is calculated.
|1. Antibody coated plate||Microtiter plate||1 plate (96 wells)||Rabbit anti-mouse /rat Ucn1 antibody coated|
|2. Standard||lyophilized||1 vial (100 ng)||Synthetic mouse/rat Ucn1|
|3. Labeled antigen||lyophilized||1 vial||Biotinylated mouse/rat Ucn1|
|4. SA-HRP solution||liquid||1 bottle (12 mL)||Horse radish peroxidase labeled streptoavidin|
|5. Enzyme substrate solution||liquid||1 bottle (12 mL)||3,3’,5,5’-Tetramethylbenzidine (TMB)|
|6. Stopping solution||liquid||1 bottle (12 mL)||1M H2SO4|
|7. Buffer solution||liquid||1 bottle (15mL)||Tris-HCl/saline buffer|
|8. Washing solution (concentrated)||liquid||1 bottle (50 mL)||Concentrated salineh|
|9. Adhesive foil||3 pieces|
Urocortin 1 EIA kit