Product Name Canine 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
Assay Range 4.9-10000pg/mL
Cross-Reactivity No cross-reactivity observed with available related molecules
Gene Names NPPA; ANF; ANP
Gene ID 608289
Swissprot ID P07499
Standard Curve Canine ANP EasyTest™ EIA Kit
• 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
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.
In blood, ANP binds with big proteins. Acidification of blood is required to separate and purify ANP.
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.
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.
Canine ANP EasyTest™ EIA Kit is for Research Use Only.
1 Plate Kit, 2 Plate Kit, 5 Plate Kit