Recommended Dilution (serum & plasma):
Human: 4X / Mouse: 4X / Rat: 4X
This kit targets the common sequence of human, mouse and rat, and thus may be used to detect ANP expression in all these species with high specificity and sensitivity.
Cross Reactivity: This EIA kit shows no cross-reactivity with any of the cytokines tested: Ghrelin, Nesfatin, Angiotensin II, NPY and APC.
Number of Targets Detected:
Compatible Sample Types:
- Cell Culture Supernatants
Method of Detection:
1, 2, or 5 x 96-Well Strip Microplate Kit
Protein Name & Synonyms:
Natriuretic peptides A (CDD-ANF) (Prepronatriodilatin) [Cleaved into: Cardiodilatin-related peptide (CDP) Atrial natriuretic factor (ANF) (Atrial natriuretic peptide) (ANP)]
Atrial natriuretic peptide (ANP) is a 28-amino acid peptide hormone secreted by cardiac myocytes of the atrium. ANP plays an important role in the homeostatic regulation of body water, sodium, potassium and fat, by acting to reduce the water, sodium and adipose loads on the circulatory system, thus reducing blood pressure.
ANP peptide contains a 17-amino acid ring which is formed by a disulfide bond between two cysteine residues at positions 7 and 23. ANP is closely related to BNP (brain natriuretic peptide) and CNP (C-type natriuretic peptide), which all share the same amino acid ring.
The mechanism of ANP-induced vasodilatation is through binding to a specific set of ANP receptors. Receptor-agonist binding causes a reduction in blood volume and therefore a reduction in cardiac output and systemic blood pressure. Lipolysis is increased and renal sodium reabsorption is decreased. The overall effect of ANP on the body is to counter increases in blood pressure and volume caused by the renin-angiotensin system.
In addition to its vasodilatation effect, ANP also serves as a adipokine. Studies have shown that ANP Increases the release of free fatty acids from adipose tissue, activates adipocyte plasma membrane NPR-A receptors, and increases intracellular cGMP levels that induce the phosphorylation of a hormone-sensitive lipase and perilipin A.
- 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
- 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 Atrial natriuretic 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.