Human BNP (Brain Natriuretic Peptide) ELISA Kit (HUES01769)
- SKU:
- HUES01769
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P16860
- Sensitivity:
- 18.75pg/mL
- Range:
- 31.25-2000pg/mL
- ELISA Type:
- Competitive
- Synonyms:
- NPPB, GC-B, B-Type Natriuretic Peptide, Natriuretic Peptide Precursor B
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Signal Transduction
Description
Human BNP (Brain Natriuretic Peptide) ELISA Kit
The Human BNP (Brain Natriuretic Peptide) ELISA Kit is a specialized assay designed for the precise measurement of BNP levels in human samples including serum, plasma, and cell culture supernatants. This kit offers exceptional accuracy and specificity, ensuring consistent and reproducible results for various research purposes. BNP is a key hormone involved in regulating blood pressure and fluid balance, particularly in the cardiovascular system.
Elevated levels of BNP are commonly associated with heart failure and other cardiac conditions, making it a valuable biomarker for diagnostic and therapeutic purposes. Researchers and healthcare professionals can utilize this ELISA kit to study BNP levels in human samples, uncovering valuable insights into cardiovascular diseases and potential treatment strategies. Trust in the reliability and performance of this kit for your research needs.
Assay type: | Competitive-ELISA |
Format: | 96T |
Assay time: | 2.5h |
Reactivity: | Human |
Detection Method: | Colormetric |
Detection Range: | 31.25-2000 pg/mL |
Sensitivity: | 18.75 pg/mL |
Sample Volume: | 50µL |
Sample Type: | Serum, plasma and other biological fluids |
Specificity: | This kit recognizes Human BNP in samples. No significant cross-reactivity or interference between Human BNP and analogues was observed. |
This ELISA kit uses Competitive-ELISA as the method. The microtiter plate provided in this kit has been pre-coated with Human BNP. During the reaction, Human BNP in the sample or standard competes with a fixed amount of Human BNP on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Human BNP. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by adding Stop Solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Human BNP in the samples is then determined by comparing the OD of the samples to the standard curve.
UniProt Protein Function: | NPPB: Cardiac hormone which may function as a paracrine antifibrotic factor in the heart. Also plays a key role in cardiovascular homeostasis through natriuresis, diuresis, vasorelaxation, and inhibition of renin and aldosterone secretion. Specifically binds and stimulates the cGMP production of the NPR1 receptor. Binds the clearance receptor NPR3. Belongs to the natriuretic peptide family. |
UniProt Protein Details: | Protein type:Secreted; Secreted, signal peptide; Hormone Chromosomal Location of Human Ortholog: 1p36. 2 Cellular Component: extracellular region; extracellular space; protein complex Molecular Function:diuretic hormone activity; hormone activity; protein binding; receptor binding Biological Process: body fluid secretion; cell surface receptor linked signal transduction; cGMP biosynthetic process; negative regulation of angiogenesis; negative regulation of cell growth; receptor guanylyl cyclase signaling pathway; regulation of blood pressure; regulation of vascular permeability; regulation of vasodilation |
NCBI Summary: | This gene is a member of the natriuretic peptide family and encodes a secreted protein which functions as a cardiac hormone. The protein undergoes two cleavage events, one within the cell and a second after secretion into the blood. The protein's biological actions include natriuresis, diuresis, vasorelaxation, inhibition of renin and aldosterone secretion, and a key role in cardiovascular homeostasis. A high concentration of this protein in the bloodstream is indicative of heart failure. The protein also acts as an antimicrobial peptide with antibacterial and antifungal activity. Mutations in this gene have been associated with postmenopausal osteoporosis. [provided by RefSeq, Nov 2014] |
UniProt Code: | P16860 |
NCBI GenInfo Identifier: | 113836 |
NCBI Gene ID: | 4879 |
NCBI Accession: | P16860. 1 |
UniProt Secondary Accession: | P16860,Q6FGY0, Q9P2Q7, B0ZBE9, |
UniProt Related Accession: | P16860 |
Molecular Weight: | 14,726 Da |
NCBI Full Name: | Natriuretic peptides B |
NCBI Synonym Full Names: | natriuretic peptide B |
NCBI Official Symbol: | NPPB |
NCBI Official Synonym Symbols: | BNP |
NCBI Protein Information: | natriuretic peptides B |
UniProt Protein Name: | Natriuretic peptides B |
UniProt Synonym Protein Names: | Gamma-brain natriuretic peptide |
Protein Family: | Brain natriuretic peptide |
UniProt Gene Name: | NPPB |
UniProt Entry Name: | ANFB_HUMAN |
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Concentration(pg/mL) | O.D | Average |
2000 | 0.292 0.334 | 0.313 |
1000 | 0.403 0.409 | 0.406 |
500 | 0.583 0.571 | 0.577 |
250 | 0.836 0.876 | 0.856 |
125 | 1.233 1.233 | 1.233 |
62.5 | 1.644 1.62 | 1.632 |
31.25 | 1.953 1.963 | 1.958 |
0 | 2.42 2.422 | 2.421 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human BNP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human BNP were tested on 3 different plates, 20 replicates in each plate.
Intra-assay Precision | Inter-assay Precision | |||||
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 20 | 20 | 20 |
Mean (pg/mL) | 100.20 | 295.40 | 980.60 | 107.90 | 303.50 | 939.20 |
Standard deviation | 6.40 | 13.90 | 49.00 | 6.80 | 12.40 | 33.80 |
C V (%) | 6.39 | 4.71 | 5.00 | 6.30 | 4.09 | 3.60 |
Recovery
The recovery of Human BNP spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type | Range (%) | Average Recovery (%) |
Serum (n=5) | 90-106 | 98 |
EDTA plasma (n=5) | 90-106 | 97 |
Cell culture media (n=5) | 92-105 | 98 |
Linearity
Samples were spiked with high concentrations of Human BNP and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
1:2 | Range (%) | 99-115 | 85-97 | 96-113 |
Average (%) | 105 | 92 | 104 | |
1:4 | Range (%) | 86-99 | 91-108 | 98-112 |
Average (%) | 93 | 98 | 104 | |
1:8 | Range (%) | 88-102 | 87-100 | 98-113 |
Average (%) | 94 | 92 | 104 | |
1:16 | Range (%) | 89-102 | 93-105 | 97-115 |
Average (%) | 96 | 100 | 105 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
Item | Specifications | Storage |
Micro ELISA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
Reference Standard | 2 vials | |
Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
HRP Conjugate Diluent | 1 vial, 14 mL | |
Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
Substrate Reagent | 1 vial, 10 mL | 4°C(shading light) |
Stop Solution | 1 vial, 10 mL | 4°C |
Plate Sealer | 5 pieces | |
Product Description | 1 copy | |
Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record their positions. It is recommended to measure each standard and sample in duplicate. Note: add all solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensure solutions do not foam when adding to the wells.
- Add 50 µL of Standard, Blank or Sample to their respective wells. The blank well is added with Sample / Standard dilution buffer.
- Immediately add 50 µL of Biotin-detection antibody working solution to each well.
- Cover with a plate seal and gently tap the plate to ensure thorough mixing. Incubate for 45minutes at 37 °C.
- Aspirate or decant the solution from the plate and add 350 µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100 µL of HRP Conjugate working solution to each well and over with a plate seal. Incubate for 30 minutes at 37 °C.
- Repeat the aspiration/wash process 5 times according to step 5.
- Add 90 µL of the Substrate reagent to each well and cover with a new plate seal. Incubatefor approximately 15 minutes at 37 °C and protect from light. The reaction time can beshortened or extended according to the colour change, but not by more than 30 minutes. Whenapparent gradient appears in standard wells, terminate the reaction.
- Stop: Add 50 µL of Stop Solution to each well (wells will develop a yellow color immediately). Note: Adding the stop solution should be done in the same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm. In advance, preheat the instrument and set the testing parameters.