useful for quantitative determination of obestatin in mouse/rat serum samples.

Mouse/Rat Obestatin EIA Kit


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Background

Obestatin is a 23 amino acid residues peptide isolated from the rat stomach. The peptide shares the precursor with a food intake stimulating peptide, ghrelin, but possesses reducing effects on food intake, gut motility and body weight (1). With the use of an antiserum directed against the mouse/rat obestatin, obestatin immunoreactivity (irOBS) was detected in cells of the gastric mucosa and myenteric plexus and in Leydig cells of the testis in Sprague–Dawley rats. Double labeling of myenteric plexus with antisera against obestatin and choline acetyltransferase (ChAT) revealed that nearly all irOBS neurons were ChAT positive and vice versa (2). Obestatin (100nM) added to dissociated and cultured rat cerebral cortical neurons elevated cytosolic calcium concentrations [Ca+2]i in a population of cortical neurons (2). Intracerebroventricular admini- stration of obestatin inhibited water drinking in ad libitum fed and watered rats, and in food and water deprived animals. In addition, obestatin inhibited angiotensin II-induced water drinking in animals provided free access to water and food (3). Obestatin peptides had no effect on insulin sensitivity as revealed by hypoglycaemic response when co-administered with insulin, supporting a role of obestatin in regulating metabolism through changes of appetite, but indicating no direct actions on glucose homeostasis or insulin secretion (4). It is supposed that in rats the effects of obestatin on food intake may be secondary to an action of the peptide to inhibit water drinking (3).

The obestatin concerning study for energy homeostasis and body weight regulation could be expected to have a large development in the future. The mouse/rat obestatin EIA assay kit developed by our laboratory can be used for direct determination of serum obestatin level’s variations and will be a useful tool for further development of obestatin research.


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Features

This mouse/rat obestatin EIA kit is used for quantitative determination of obestatin in mouse/rat serum samples.
It has various advantages, such as highly specific and sensitive quantification, no influences with other body fluids or physiological active substances and unnecessity of sample pretreatment. Mouse/rat obestatin standard of this kit is a highly purified synthetic product (purity: higher than 99%).

  • The assay kit can measure mouse/rat obestatin within the range of 0.082-20ng/mL.
  • The assay completes within 18-20h. + 1.5 h.
  • With one assay kit, 41 samples can be measured in duplicate.
  • Test sample: mouse or rat serum. Sample volume: 25 μL.
  • The 96-well plate of this kit is consists of 8-well strips, so that divided use by strips is possible at user’s option.
  • Precision and reproducibility
    Intra-assay CVi“j3.7`6.9 (mouse serum), 3.4`6.7 (rat serum).
    Inter-assay CVi“j4.5`8.4 (mouse serum), 8.1`10.8 (rat serum).
  • Precision and reproducibility
    Intra-assay CVi“j3.7`6.9 (mouse serum), 3.4`6.7 (rat serum).
    Inter-assay CVi“j4.5`8.4 (mouse serum), 8.1`10.8 (rat serum).
  • 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 shows cross-reactivity of 100% to mouse/rat obestatin, 118.6% to mouse/rat obestatin (11-23)-NH2, 0.5% to mouse/rat obestatin (1-23)-OH and less than 0.39% to human/ mouse/rat obestatin (1-10) and no cross-reactivity to human obestatin, human obestatin (11- 23))-NH2, and mouse/rat ghrelin and mouse/rat des-octanoyl ghrelin.

<Test Principle>
This EIA kit for determination of obestatin in mouse/rat serum samples is based on a competitive enzyme immunoassay using the combination of highly specific antibody to mouse/ rat obestatin@and biotin–avidin affinity system. The 96 wells plate is coated with goat anti rabbit IgG, to which biotinylated mouse/rat obestatin, mouse/rat obestatin standard or samples and rabbit anti mouse/rat obestatin antibody are added for competitive immunoreaction. After incubation and plate washing, horse radish peroxidase (HRP) labeled streptavidin (SA) is added, so that HRP labeled SA-biotinylated mouse/rat obestatin-antibody complex is formed on the surface of the wells. Finally, HRP enzyme activity is determined by 3,3’,5,5’-Tetramethylbenzidine (TMB) and the concentration of mouse/rat obestatin is calculated.

Composition

Component Form Quantity Main@Ingredient
1. Antibody coated plate microtiter plate 1 plate
(96 wells)
Goat anti rabbit IgG
2. Standard Lyophilized powder 1 vial
(20ng)
Synthetic mouse/rat obestatin
3. Labeled antigen Lyophilized powder 1 vial Biotinylated mouse/rat obestatin
4. Specific antibody liquid 1 bottle
(6 mL)
Rabbit anti mouse/rat obestatinAntibody
5. SA-HRP solution liquid 1 bottle
(12 mL)
HRP labeled streptoavidin
6. TMB substrate liquid 1 bottle
(12 mL)
3,3’,5,5’-Tetramethylbenzidine (TMB)
7. Reaction Stopping solution liquid 1 bottle
(12 mL)
1M H2SO4
8. Buffer solution liquid 1 bottle
(25 mL)
BSA containing saline buffer
9. Washing solution
     (concentrated)
liquid 1 bottle
(25 mL)
Concentrated saline
10.Adhesive foil   3 sheets  
Product List

Product Name Cat# Quantity Price

Mouse/Rat Obestatin, EIA Kit DataSheet ,

YII-YK230-EX

1 KIT ¥76,000
$1014

Related Products [ antibody ]


Product Name Cat# Quantity Price

Anti Obestatin, Rat (Rabbit) Unlabeled DataSheet

YII-Y460-EX

50 UL ¥35,000
$467

Antigen : Obestatin (Mouse, Rat)
IHC : 1 : 5,000
EIA : 1 : 27,000
Specificity :
Based on non-competitive EIA:Obestatin (mouse, rat) 100%,
Obestatin (11-23) (mouse, rat) 90.42%, Obestatin (1-10)(mouse, rat, human) 25.95%,
Obestatin (1-23)-OH (mouse, rat) 20.76%, Obestatin (human) 26.37%,
Obestatin (human) (11-23) 24.91%, Ghrelin (mouse, rat) 0%, Des-octanonyl ghrelin (mouse, rat) 0%


“Mouse/Rat Obestatin EIA Kit” also belongs to the following categories.