Equine ANP EasyTest™ EIA Kit

Product Features

Product Name            Equine ANP EasyTest™ EIA Kit
Assay Type                   Solid Phase Competitive ELISA
Format                           96-well strip plate
Assay Process             Two wash step, three incubations
Assay Length              2.0 hours
Sample Volume         Serum , Plasma, 50µL
Detection Method    Colorimetric
Sensitivity                   15pg/mL
Assay Range                4.9-10000pg/mL
Reactivity                     Equine
Cross-Reactivity       No cross-reactivity observed with available related molecules
Gene Names               NPPA
Gene ID                         100034201
Swissprot ID               P27104

Standard Curve        Equine ANP EasyTest™ EIA Kit

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Anti-ANP Antibody
• Standard ANP peptide
• Biotinylated ANP peptide
• TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 25 µl of anti-ANP antibody to all wells.
3. Add 75 µl of 1 x assay diluent to the blank wells and 50 µl to B0 wells.
4. Add 50 µl of standard peptide/samples to the appropriate wells.
5. Add 25 µl of biotinylated peptide into each well except the blank wells.
6. Incubate 1.0 h at RT.
7. Wash and add 100 µl of diluted streptavidin-HRP solution to all wells.
8. Incubate 45 min at RT.
9. Wash and add 100 µl of TMB Substrate Solution into each well.
10. Incubate 15 min at RT.
11. Add 50 µl of Stop Solution to each well.
12. Read at 450 nm/570 nm.

Application

In blood, ANP binds with big proteins. Acidification of blood is required to separate and purify ANP.

Background

Atrial natriuretic peptide (ANP), atrial natriuretic factor (ANF), is a powerful vasodilator, and a polypeptide hormone secreted by heart muscle cells. ANP is a 28-amino acid peptide with a 17-amino acid ring in the middle of the molecule.

ANP is secreted in response to stretching of the atrial wall，reduced Sympathetic stimulation of βadrenoceptors, hypernatremia, Angiotensin-II, Endothelin and exercise. It is released by muscle cells in the atria of the heart in response to high blood volume. ANP acts to reduce the water, sodium and adipose loads on the circulatory system, thereby reducing blood pressure. ANP has exactly the opposite function of the aldosterone secreted by the zona glomerulosa in regard to its effect on sodium in the kidney.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status      

Equine ANP EasyTest™ EIA Kit is for Research Use Only.

• Manual
• AssayProcedure
• MSDS