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Antigen Information

Accession Number:

P16860

Gene ID:

4879

Gene Symbols:

NPPB

Protein Name & Synonyms:

Natriuretic peptides B (Gamma-brain natriuretic peptide) [Cleaved into: Brain natriuretic peptide 32 (BNP(1-32)) (BNP-32) BNP(1-30) BNP(1-29) BNP(1-28) BNP(2-31) BNP(3-32) BNP(3-30) BNP(3-29) BNP(4-32) BNP(4-31) BNP(4-30) BNP(4-29) BNP(4-27) BNP(5-32) BNP(5-31) BNP(5-29)]

Assay Format

Detection Range:

0.1-1,000 pg/ml

Recommended Dilution (serum & plasma):

Human: 2X

Sensitivity:

1.66 pg/ml

Specificity:

Cross Reactivity: This EIA kit shows no cross-reactivity with any of the adipokines tested: Ghrelin, Nesfatin, Angiotensin II, NPY and APC.

Number of Targets Detected:

Species Detected:

Human

Compatible Sample Types:

Cell Culture Supernatants
Serum

Design Principle:

Competition-based

Method of Detection:

Colorimetric

Quantitative/Semi-Quantitative:

Quantitative

Solid Support:

96-well Microplate

Product Specifications

Size:

1, 2, or 5 x 96-Well Strip Microplate Kit

Introduction

Brain natriuretic peptide (BNP), (aka B-type natriuretic peptide), is a 32 amino acid polypeptide secreted by the ventricles of the heart in response to excessive stretching of myocytes in the ventricles. BNP was originally identified in extracts of porcine brain, but in humans it is produced mainly in the cardiac ventricles. Its counterpart in rats is a 45 amino acid peptide hormone. At the time of release, a co-secreted 76 amino acid N-terminal fragment (NT-proBNP) is also released with BNP.

BNP binds to and activates NPRA in a similar fashion to atrial natriuretic peptide (ANP) but with 10-fold lower affinity. The biological half-life of BNP, however, is twice as long as that of ANP. Both ANP and BNP have limited ability to bind and activate NPRB.

Physiologic actions of BNP include decrease in systemic vascular resistance and central venous pressure as well as an increase in natriuresis. Thus, the resulting effect of BNP is a decrease in cardiac output and a decrease in blood volume.

Tests showing elevated levels of BNP or NT-proBNP in blood are used as a diagnosis of heart failure and may be useful to establish prognosis in heart failure, as both markers are typically higher in patients with poorer outcome.

Both BNP and NT-proBNP have been approved as a marker for acute congestive heart failure (CHF). The plasma concentrations of both BNP are increased in patients with asymptomatic and symptomatic left ventricular dysfunction. There is no level of BNP that perfectly separates patients with and without heart failure.

BNP EIA/ELISA Test Kit Features

Strip plates and additional reagents allow for use in multiple experiments
Quantitative protein detection
Establishes normal range
The best products for confirmation of antibody array data

Application Notes

Pre-Coated 96-well Strip Microplate
Wash Buffer
Standard Peptide
Assay Diluent(s)
Biotinylated Peptide
HRP-Streptavidin
TMB One-Step Substrate
Stop Solution
Assay Diagram
Positive Control Sample
Capture Antibody
User Manual

Other Materials Required

Distilled or deionized water
Precision pipettes to deliver 2 µl to 1 ml volumes
Adjustable 1-25 ml pipettes for reagent preparation
100 ml and 1 liter graduated cylinders
Tubes to prepare standard and sample dilutions
Orbital shaker
Aluminum foil
Saran Wrap
Absorbent paper
Microplate reader capable of measuring absorbance at 450nm
SigmaPlot software (or other software that can perform four-parameter logistic regression models)

Protocol Outline

Prepare all reagents, samples and standards as instructed.
Add 100 µl detection antibody to each well.
Incubate 1.5 h at RT or O/N at 4°C.
Add 100 µl standard or sample to each well.
Incubate 2.5 h at RT.
Add 100 µl prepared streptavidin solution.
Incubate 45 min at RT.
Add 100 µl TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 µl Stop Solution to each well.
Read plate at 450 nm immediately.

Storage/Stability

Standard, Biotinylated Brain Natriuretic peptide, and Positive Control should be stored at -20°C after arrival. Avoid multiple freeze-thaws. The remaining kit components may be stored at 4°C. Opened Microplate Wells and antibody (Item N) may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge.