Human NT-ProANP Quantikine ELISA Kit Summary

Product Summary

The Quantikine^® Human NT-ProANP Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human NT-ProANP in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human NT-ProANP and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human NT-ProANP showed linear curves that were parallel to the standard curves obtained using the Quantikine® kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human NT-ProANP.

Preparation and Storage

Background:  Atrial Natriuretic Peptide/ANP

Atrial Natriuretic peptide (ANP) is synthesized as a 151 aa preproprotein. The proprotein contains cardiodilatin (aa 26 - 92) and atrial natriuretic factor (ANF; aa 124 - 151) which become active after processing by the serine protease corin. ANP is primarily synthesized in the heart and is secreted in response to atrial distention. ANF circulates in the plasma, eliciting natriuretic, diuretic, vasorelaxant and antimitogenic effects directed toward reduction of body fluid and blood pressure regulation. ANF acts through the cGMP-coupled NP receptor A. Human ANP and ANF share 84% and 96% aa identity with mouse ANP and ANF, respectively.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1、Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2、Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3、Add 50 μL of Assay Diluent to each well.

4、Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5、Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6、Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7、Aspirate and wash 5 times.

8、Add 100 μL Substrate Solution to each well.

9、Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.