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Assay Format
Detection Range:
0.1-1,000 ng/ml
Recommended Dilution (serum & plasma):
Human: 2X / Mouse: 2X / Rat: 2X
Sensitivity:
15.8 ng/ml
Specificity:
This kit is designed to target the C-terminus of the 77-aa apelin peptide and therefore is expected to detect all active forms of Apelin including Apelin-36, Apelin-31, Apelin-28 and Apelin 13 can be detected.
Cross Reactivity: This kit shows no cross-reactivity with any of the cytokines tested: Ghrelin, Nesfatin, and NPY.
Cross Reactivity: This kit shows no cross-reactivity with any of the cytokines tested: Ghrelin, Nesfatin, and NPY.
Number of Targets Detected:
1
Species Detected:
- Human
- Mouse
- Rat
Compatible Sample Types:
- Cell Culture Supernatants
- Serum
Design Principle:
- Competition-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
- Quantitative
Solid Support:
96-well Microplate
Product Specifications
Size:
1, 2, or 5 x 96-Well Strip Microplate Kit
Protein Information
Accession Number:
- Q9ULZ1
Gene ID:
- 8862
Gene Symbols:
- APEL
- APLN
Protein Name & Synonyms:
Apelin (APJ endogenous ligand) [Cleaved into: Apelin-36 Apelin-31 Apelin-28 Apelin-13]
Introduction
Apelin, an endogenous ligand for the G-protein-coupled APJ receptor, has been recently extensively studied in obesity research. It is not only expressed in adipocyte tissue, but also widely expressed in various other organs such as the heart, lung, kidney, gastrointestinal tract, brain, adrenal glands, endothelium, and human plasma.
Apelin is derived from a 77-amino-acid prepropeptide that is cleaved into a 55-amino-acid fragment and then into shorter forms. The physiologically active form is thought to be apelin 36, although the pyroglutamylated form of apelin 13, which is also produced endogenously, is more potent.
Studies have shown the association between apelin and obesity. Apelin has higher circulating levels in obesity. Insulin exerts a positive action on adipocyte apelin production. Apelin also regulates fluid homeostasis, playing an important role in the hypothalamic regulation of food and water intake, and pituitary hormone release.
In addition to its role in obesity, apelin acts as a mediator of cardiovascular control, including for blood pressure and blood flow. It is one of the most potent stimulators of cardiac contractility yet identified, and plays a role in cardiac tissue remodeling. Apelin levels are increased in left ventricles of patients with chronic heart failure and also in patients with chronic liver disease.
Apelin is derived from a 77-amino-acid prepropeptide that is cleaved into a 55-amino-acid fragment and then into shorter forms. The physiologically active form is thought to be apelin 36, although the pyroglutamylated form of apelin 13, which is also produced endogenously, is more potent.
Studies have shown the association between apelin and obesity. Apelin has higher circulating levels in obesity. Insulin exerts a positive action on adipocyte apelin production. Apelin also regulates fluid homeostasis, playing an important role in the hypothalamic regulation of food and water intake, and pituitary hormone release.
In addition to its role in obesity, apelin acts as a mediator of cardiovascular control, including for blood pressure and blood flow. It is one of the most potent stimulators of cardiac contractility yet identified, and plays a role in cardiac tissue remodeling. Apelin levels are increased in left ventricles of patients with chronic heart failure and also in patients with chronic liver disease.
Product Features
- 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
Application Notes
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Standard Peptide
- Assay Diluent(s)
- Biotinylated Peptide
- HRP-Streptavidin
- 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 Outline
- Prepare 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.
Storage/Stability
Standard, Biotinylated Apelin C-Terminus 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.
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Najafipour H., Vakili A., Shahouzehi B., et al. Investigation of changes in apelin receptor mRNA and protein expression in the myocardium and aorta of rats with two-kidney, one-clip (2K1C) Goldblatt hypertension. J Physiol Biochem. 2015 Jun;71(2):165-75. doi: 10.1007/s13105-015-0394-z
Species: Rat
Sample type: Serum -
Sanchis-Gomar F., Alis R., Rampinini E., et al. Adropin and apelin fluctuations throughout a season in professional soccer players: Are they related with performance? Peptides. 2015 Aug;70:32-6. doi: 10.1016/j.peptides.2015.05.001
Species: Human
Sample type: Serum -
Gileles-Hillel A., Alonso-Alvares M., Kheirandish-Gozal L., Peris E., et al. Inflammatory markers and obstructive sleep apnea in obese children: The NANOS Study. Mediators of Inflammation. Accepted May 2014 http://www.hindawi.com/journals/mi/aip/605280/ [Epub ahead of print]
Species: Human
Sample type: Plasma -
Samy D., Ismail C., Deif A., et al. Induction of Apelin by Losartan in Renal Ischemia/Reperfusion Injury in Rats - Implication of Endothelial Nitric Oxide Synthase (eNOS) Phosphorylation. J Phys Pharm Adv 2014 , 4 (11): 465 - 477 DOI: 10.5455/jppa.20141117043349
Species: Rat
Sample type: Tissue Lysate -
Than A., Cheng Y., Foh LC., et al. Apelin inhibits adipogenesis and lipolysis through distinct molecular pathways. Mol Cell Endocrinol. 2012 Oct 15;362(1-2):227-41. doi: 10.1016/j.mce.2012.07.002.
Species: Mouse
Sample type: Conditioned Media -
Than A., Tee W., Chen P. Apelin secretion and expression of apelin receptors in 3T3-L1 adipocytes are differentially regulated by angiotensin type 1 and type 2 receptors. Mol Cell Endocrinol. 2012 Apr 4;351(2):296-305. doi: 10.1016/j.mce.2012.01.005
Species: Mouse
Sample type: Conditioned Media -
Najafipour H., Soltani Hekmat A., Nekooian A., Esmaeili-Mahini S. Apelin receptor expression in ischemic and non- ischemic kidneys and cardiovascular responses to apelin in chronic two-kidney-one-clip hypertension in rats. Regul Pept. 2012 Oct 10;178(1-3):43-50. doi: 10.1016/j.regpep.2012.06.006.
Species: Rat
Sample type: Plasma -
Geiger, K., et al. "Hypoxia induces apelin expression in human adipocytes." Hormone and metabolic research 43.06 (2011): 380-385.
Species: Human
Sample type: Conditioned Media
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Usage
This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.
