Obestatin is a 24-aa peptide hormone that is produced in the stomach and small intestine of humans and several other mammals. It is encoded by the same gene that also encodes adipokine Ghrelin, which is initially expressed as a preprohormone that is subsequently cleaved into two smaller peptides, Ghrelin and Obestatin. A model has been proposed suggesting a "Yin-Yang" relationship between the two peptides, that Ghrelin and Obestatin have countering effects to each other.
There are reports using Ghrelin knockout mice studies which showed that removing the Ghrelin gene from mice did not significantly reduce their appetite. Additional reports show that Obestatin antagonizes growth hormone secretion and food intake induced by Ghrelin.
Obestatin has shown some potential clinical applications. Studies on the Obestatin/Ghrelin ratio in the gastrointestinal tract and plasma are associated with metabolic diseases such as irritable bowel syndrome, obesity and type II diabetes mellitus.
- Strip plates and additional reagents allow for use in multiple experiments
- Quantitative protein detection
- Establishes normal range
- The best products for confirmation of antibody array data
The capture antibody provided in this kit recognizes human, mouse and rat Obestatin.
Target Protein Information
Appetite-regulating hormone (Growth hormone secretagogue) (Growth hormone-releasing peptide) (Motilin-related peptide) (Protein M46) [Cleaved into: Obestatin]
GHRL MTLRP UNQ524/PRO1066
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Standard Peptide
- Assay Diluent(s)
- Biotinylated Peptide
- TMB One-Step Substrate
- Stop Solution
- Assay Diagram
- Positive Control Sample
- Capture Antibody
- User Manual
Other Materials Required
- Distilled or deionized water
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare standard and sample dilutions
- Orbital shaker
- Aluminum foil
- Saran Wrap
- Absorbent paper
- Microplate reader capable of measuring absorbance at 450nm
- SigmaPlot software (or other software that can perform four-parameter logistic regression models)
Protocol OutlinePrepare all reagents, samples and standards as instructed.Add 100 µl detection antibody to each well.Incubate 1.5 h at RT or O/N at 4°C.Add 100 µl standard or sample to each well.Incubate 2.5 h at RT.Add 100 µl prepared streptavidin solution.Incubate 45 min at RT.Add 100 µl TMB One-Step Substrate Reagent to each well.Incubate 30 min at RT.Add 50 µl Stop Solution to each well.Read plate at 450 nm immediately.
Standard, Biotinylated Obestatin peptide, and Positive Control should be stored at -20°C after arrival. Avoid multiple freeze-thaws. The remaining kit components may be stored at 4°C. Opened Microplate Wells and antibody (Item N) may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge.