NT-proCNP ELISA Assay Kit

$630.00

The NT-proCNP ELISA Assay KIT(enzyme-linked immunoassay kit) is intended for the quantitative determination of human NT proCNP in plasma and serum. The Eagle Biosciences NT-proCNP ELISA Assay KIT is manufactured by Biomedica (www.bmgrp.com).  The Eagle Biosciences proCNP ELISA Assay KIT is for research use only and not to be used in diagnostic procedures.

SKU: BI-20812 Categories: ,

NT-proCNP ELISA Assay Kit

NT-proCNP ELISA Assay Kit Developed and Manufactured in Austria by Biomedica

Size: 1×96 wells
Sensitivity: 0.7 pmol
Dynamic Range: 0-128 pmol/L
Incubation Time: 4 hours
Sample Type: Serum, EDTA plasma, heparin plasma, and citrate plasma
Sample Size: 20 µL
Alternative Names: C–type natriuretic peptide
For Research Use Only

Controls Included


Conversion factor: 1pg/ml = 0.20 pmol/l
Reference Values:
Median serum (n=32) = 14.5 pmol/l
Median EDTA plasma (n=33) = 15 pmol/l
Median heparin plasma (n=18) = 13.5 pmol/l
Median citrate plasma (n=18) = 12 pmol/l
Each laboratory should establish its own reference range for the samples under investigation. Do not change sample type during the study.


Assay Principle

The NT-proCNP ELISA Assay Kit is a sandwich enzyme immunoassay for the quantitative determination of NT-proCNP in human serum, plasma, urine, cell culture supernatants and non-human samples (rat, mouse, pig). In a first step, assay buffer and standard/control/sample are pipetted into the wells of the microtiter strips, which are pre-coated with polyclonal sheep anti-NT-proCNP antibody, for a short incubation. Without the need of a washing step, conjugate (sheep anti-human NT-proCNP-HRP) is added into the wells. NT-proCNP present in the standrad/sample/control binds to the precoated antibody in the well and forms a sandwich with the detection antibody. In the washing step all non-specific unbound material is removed. After a final washing step, the substrate (TMB, tetramethylbenzidine) is pipetted into the wells. The enzyme-catalyzed color change of the substrate is directly proportional to the amount of NT-proCNP present in the sample. This color change is detectable with a standard microtiter plate ELISA reader.

The NT-proCNP ELISA kit uses highly purified, epitope-mapped antibodies. The antibodies utilized in the NT-proCNP ELISA (BI-20892) are as follows:

– Capture antibody: polyclonal sheep anti-NT-proCNP (epitope: AA62-68 of Uniprot ID P23582)

– Detection antibody: polyclonal sheep anti-NT-proANP (epitope: AA34-37 of Uniprot ID P23582)


Related Products

NT-proBNP ELISA Assay Kit
proANP ELISA Assay Kit
BNP Fragment (Nt-proBNP 8-29) ELISA Assay Kit

Learn more about CNP at Eagle Biosciences Biomarker Spotlight page dedicated to CNP here:
EagleBio Spotlight: CNP

Additional Information

Assay Background


CNP belongs to a well characterised group of natriuretic peptides like atrial natriuretic (ANP) and brain natriuretic peptide (BNP), which play various important roles as regulatory mechanisms as well as bio-markers. They participate in the regulation of blood pressure and body fluid homeostasis, modify growth and development of cardiovascular tissues and bone. ANP and BNP are cardiac hormones, secreted by the atria and ventricles, respectively, in the normal adult heart. In contrast to ANP and BNP, very little CNP is produced by cardiac tissue, but is expressed primarily in the brain, pituitary gland, vascular endothelium, kidney and female reproductive tract.

All natriuretic peptides are produced as propeptides, which are subsequently cleaved into the biologically active, C-terminal hormone and the N-terminal fragment (NT-proANP 1-98, NT-proBNP 1-76 and NT-proCNP, which length is still under discussion). Since those N-terminal fragments are much more stable, and circulate in higher amounts than the active hormones, they are easier and more reliable to be measured in serum or plasma. So we developed a sandwich immunoassay for NT-proCNP using antibodies directed against amino acids 1-19 and 30-50. The physiological role of CNP is not only studied in cardiac disease, but also in bone developmental biology, generally in bone research, renal diseases, embryonic developmental research and vascular diseases.

Assay Principle


  1. All reagents and samples have to be brought to room temperature (18-26°C) before they can be used in the NT-proCNP ELISA Assay Kit.
  2. Mark position for BLANK/STD/SAMPLE/CTRL (Blank/Standard/Sample/Control) on the protocol sheet.
  3. Take microtiter strips out of the aluminium bag, reserve a minimum of one well as blank. Unused strips can be stored with desiccant in the aluminium bag at +4°C (2-8°C) until the expiry date.
  4. Add 50 µl STD/SAMPLE/CTRL (Standard/Sample/Control) in duplicate into respective wells, except blank.
  5. Add 200 µl CONJ (anti NT-proCNP-HRPO) into each well, except blank, swirl gently.
  6. Cover tightly and incubate for 4 hours at room temperature (18-26°C) in the dark.
  7. Aspirate and wash wells 5x with 300 µl diluted WASHBUF (Wash buffer), remove remaining WASHBUF by hitting plate against paper towel after the final washing step.
  8. Add 200 µl SUB (Substrate) into each well.
  9. Incubate for 30 min at room temperature (18-26°C) in the dark.
  10. Add 50 µl STOP (Stop solution) into each well.
  11. Measure absorbance immediately at 450 nm with reference 630 nm, if available.

Standard Curve


nt-procnp-elisa-standard-curve

Manual

Product Manual


Publications

Citations


  • “Identification of amino-terminal pro-C-type natriuretic peptide in human plasma”  T.C.R Prickett et al., Biochem. and Biophy. Res. Comm., 286, 513-517 (2001)
  • “Overexpression of CNP in chondrocytes rescues achondroplasia through a MAPK- dependent pathway”  Akihiro Yasoda et al., Nature Medicine, 10, 80-86 (Dez 2003)
  • “C-type natriuretic peptide in reproduction, pregnancy, and fetal development”  T. Walther et al. J. of Endo, 180,17-22 (2004)

Related Resources Citing EagleBio Kits:


Stanek, B. et al. “Prognostic evaluation of neurohumoral plasma levels before and during beta-blocker therapy in advanced left ventricular dysfunction” Journal of American Cardiology 2001; 38: 436-442.

Related Kits:


proANP ELISA Assay Kit

BNP Fragment (Nt-proBNP 8-29) ELISA Assay Kit

Cardiovascular Assay Kits

Related News:


EagleBio Spotlight: ANP

EagleBio Spotlight: BNP

Interesting Study Highlighting Eagle Biosciences’ BNP Fragment ELISA Assay Kit