NPPA ELISA Kit (Mouse)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level N-Terminal Pro-Atrial Natriuretic Peptide (NT-ProANP) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level N-Terminal Pro-Atrial Natriuretic Peptide (NT-ProANP) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

A; Anf; ANP; atrial natriuretic factor prohormone; atrial natriuretic peptide prohormone; atriopeptigen; cardiodilatin; CDD; natriuretic peptide precursor type A; natriuretic peptides A; P; Pnd; preproANF; preproANP; preproCDD-ANF; prepronatriodilatin; proANF; proANP.

Hormone playing a key role in cardiovascular homeostasis through regulation of natriuresis, diuresis, and vasodilation. Also plays a role in female pregnancy by promoting trophoblast invasion and spiral artery remodeling in uterus. Specifically binds and stimulates the cGMP production of the NPR1 receptor. Binds the clearance receptor NPR3.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to N-Terminal Pro-Atrial Natriuretic Peptide (NT-ProANP) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to N-Terminal Pro-Atrial Natriuretic Peptide (NT-ProANP) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain N-Terminal Pro-Atrial Natriuretic Peptide (NT-ProANP), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of N-Terminal Pro-Atrial Natriuretic Peptide (NT-ProANP) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) serum (n=5) 98-105 101 EDTA plasma (n=5) 95-103 98 heparin plasma (n=5) 78-96 89 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 98-105 101 serum (n=5) 98-105 101 EDTA plasma (n=5) 95-103 98 EDTA plasma (n=5) 95-103 98 heparin plasma (n=5) 78-96 89 heparin plasma (n=5) 78-96 89

Component Amount Anti-Nppa Microplate 96 Wells (12 x 8 Well Strips) Nppa Lyophilized Standard 2 x 5 ng 100X Biotinylated Nppa Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-Nppa Microplate 96 Wells (12 x 8 Well Strips) Anti-Nppa Microplate 96 Wells (12 x 8 Well Strips) Nppa Lyophilized Standard 2 x 5 ng Nppa Lyophilized Standard 2 x 5 ng 100X Biotinylated Nppa Detector Antibody 120 uL 100X Biotinylated Nppa Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of N-Terminal Pro-Atrial Natriuretic Peptide (NT-ProANP) . No significant cross-reactivity or interference between N-Terminal Pro-Atrial Natriuretic Peptide (NT-ProANP) and analogues was observed.