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useful for quantitative determination of obestatin in human plasma and serum samples

Human Obestatin EIA Kit

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 SpragueDawley 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 human obestatin EIA assay kit can be used for direct determination of blood obestatin level’s variations and will be a useful tool for further development of obestatin research.

Feature and Advantages

This EIA kit is used for quantitative determination of obestatin in human plasma and 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. Human 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 0.412-100ng/mL.
  • The assay completes within 18-20h. + 1.5 h.
  • With one assay kit, 41 samples can be measured in duplicate.
  • Test sample: human plasma or serum Sample volume : 20 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(%)3.5-9.9.
      Inter-assay CV(%)5.6-9.0.
  • Stability and Storage
      Store all the components at 2-8°C.
      The kit is stable under the condition for 6 months from the date of manufacturing.

<Specificity>
The EIA kit shows cross-reactivity of 100% to human obestatin, 37.3% to mouse/rat obestatin, 25.2% to human obestatin (11-23)-NH2, less than 0.02% to human/mouse/rat obestatin (1-10), and no cross-reactivity to mouse/rat obestatin (11-23)-NH2. It shows no cross-reactivity to human ghrelin and human des-octanoyl ghrelin in the range of standard concentrations.

<Assay principle>
This EIA kit for determination of obestatin in human plasma or serum samples is based on a competitive enzyme immunoassay using the combination of highly specific antibody to human obestatin and biotinavidin affinity system. The 96 wells plate is coated with goat anti rabbit IgG, to which biotinylated human obestatin, human obestatin standard or samples and rabbit anti human 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 human 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 human 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 (50ng) Synthetic human obestatin
3. Labeled antigen Lyophilized powder 1 vial Biotinylated human obestatin
4. Specific antibody liquid 1 bottle (6 mL) Rabbit anti human 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

Human Obestatin EIA kit

YII-YK231-EX 1KIT

¥ 76,000
$ 1014
€ 760