Mouse NT proBNP ELISA Kit
- SKU:
- MOFI01007
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Sensitivity:
- 9.375pg/ml
- Range:
- 15.625-1000pg/ml
- ELISA Type:
- Sandwich
- Synonyms:
- NT-proBNP, N-Terminal Pro-Brain Natriuretic Peptide
- Reactivity:
- Mouse
Description
Mouse NT proBNP ELISA Kit
The Mouse NT-proBNP ELISA Kit is specifically designed for the quantitative measurement of N-terminal pro-brain natriuretic peptide (NT-proBNP) levels in mouse serum, plasma, and tissue lysates. NT-proBNP is a valuable biomarker for various cardiac conditions, particularly heart failure, in mice. This kit allows researchers to accurately assess NT-proBNP levels, aiding in the diagnosis, prognosis, and monitoring of cardiac diseases in mouse models.
Key Features
Save Time | Pre-coated 96 well plate | |
Quick Start | Kit includes all necessary reagents | |
Publication Ready | Reproducible and reliable results |
Overview
Product Name: | Mouse NT proBNP ELISA Kit |
Product Code: | MOFI01007 |
Size: | 96 Assays |
Alias: | NT-proBNP, N-Terminal Pro-Brain Natriuretic Peptide |
Detection Method: | Sandwich ELISA |
Application: | This immunoassay kit allows for the in vitro quantitative determination of Mouse NT-proBNP concentrations in serum plasma and other biological fluids. |
Sensitivity: | 9.375pg/ml |
Range: | 15.625-1000pg/ml |
Storage: | 4°C for 6 months |
Note: | For Research Use Only |
Additional Information
Recovery | Matrices listed below were spiked with certain level of Mouse NT-proBNP and the recovery rates were calculated by comparing the measured value to the expected amount of Mouse NT-proBNP in samples.
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Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mouse NT-proBNP and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
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CV(%) | Intra Assay <8 Inter Assay <10 |
Kit Components
Component | Quantity | Storage |
ELISA Microplate (Dismountable) | 8x12 strips | 4°C for 6 months |
Lyophilized Standard | 2 | 4°C/ -20°C |
Sample/Standard Dlution Buffer | 20ml | 4°C |
Biotin-labeled Antibody (Concentrated) | 120ul | 4°C (Protection from light) |
Antibody Dilution Buffer | 10ml | 4°C |
HRP-Streptavidin Conjugate (SABC) | 120ul | 4°C (Protect from light) |
SABC Dilution Buffer | 10ml | 4°C |
TMB Substrate | 10ml | 4°C (Protection from light) |
Stop Solution | 10ml | 4°C |
Wash Buffer (25X) | 30ml | 4°C |
Plate Sealer | 5 | - |
Other materials required:
- Microplate reader with 450 nm wavelength filter
- Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
- Incubator
- Deionized or distilled water
- Absorbent paper
- Buffer resevoir
Protocol
*Note: Protocols are specific to each batch/lot. For the exact instructions please follow the protocol included in your kit.
Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.
Step | Procedure |
1. | Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells! |
2. | Aliquot 0.1ml standard solutions into the standard wells. |
3. | Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well. |
4. | Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells. |
5. | Seal the plate with a cover and incubate at 37 °C for 90 min. |
6. | Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2. |
7. | Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall. |
8. | Seal the plate with a cover and incubate at 37°C for 60 min. |
9. | Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash. |
10. | Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min. |
11. | Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min. |
12. | Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color. |
13. | Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. |
14. | Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution. |
Sample Preparation
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
Sample Type | Protocol |
Serum | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. |
Plasma | Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit. |
Urine & Cerebrospinal Fluid | Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. |
Cell culture supernatant | Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. |
Cell lysates | Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
Tissue homogenates | The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C. |
Tissue lysates | Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C |
Breast Milk | Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. |
NT proBNP Background
N-terminal pro-brain natriuretic peptide (NT-proBNP)
What is NT-proBNP?
N-terminal pro-brain natriuretic peptide (NT-proBNP) is a protein that serves as a biomarker for the diagnosis, prognosis, and management of heart-related conditions, particularly heart failure. It is derived from the precursor peptide called pro-brain natriuretic peptide (proBNP).
NT-proBNP Gene
In mice, the gene responsible for NT-proBNP production is called Nppb, located on chromosome 4. It consists of two exons and one intron and spans approximately 4 kilobases.
The protein is initially synthesized as a larger precursor and undergoes processing to produce the active mature peptide. This mature peptide functions as a cardiac hormone, contributing to the remodeling of the heart's ventricles and regulating blood pressure. In mice, the absence of this gene leads to the development of cardiac fibrosis. In humans, variations in this gene have been linked to congestive heart failure, reduced bone-mineral density, and postmenopausal osteoporosis. Through alternative splicing, the gene produces various transcript variants that encode different isoforms of the protein.
NT-proBNP Structure
NT-proBNP in mice is a peptide derived from the larger precursor peptide, pro-brain natriuretic peptide (proBNP). Similar to humans, proBNP undergoes processing to generate the biologically inactive proBNP molecule, which is further cleaved to yield the active B-type natriuretic peptide (BNP) and the inactive NT-proBNP. NT-proBNP in mice corresponds to the N-terminal fragment of proBNP, encompassing the first 121 amino acids of the precursor molecule.
Predicted Structure of Mouse NT-proBNP. Source: Uniprot
NT-proBNP Function
NT-proBNP in mice serves as a cardiac hormone and plays a crucial role in regulating blood volume and blood pressure. It functions by promoting vasodilation, natriuresis, and diuresis, thereby reducing the workload on the heart and counteracting fluid retention. These actions help maintain cardiovascular homeostasis.
NT-proBNP Clinical Significance
NT-proBNP is widely used as a biomarker for the diagnosis, prognosis, and management of heart-related conditions in mice, including heart failure.
NT-proBNP Levels
Elevated levels of NT-proBNP in mouse models are indicative of increased cardiac stress and impaired cardiac function, similar to observations in humans. Measurement of NT-proBNP levels aids in the differentiation of heart failure from other causes of cardiac dysfunction, as well as monitoring disease progression and response to interventions in mice.
Moreover, NT-proBNP is also utilized in assessing the risk stratification and prognosis of cardiovascular events in mice. Higher NT-proBNP levels are associated with an increased likelihood of adverse outcomes, such as mortality, in both acute and chronic heart failure models. It is also instrumental in guiding treatment decisions and optimizing therapeutic strategies in mouse models of heart failure.
Mouse NT proBNP ELISA Kit FAQs
Q: What is the purpose of the Mouse NT-proBNP ELISA Kit?
The Mouse NT-proBNP ELISA Kit is specifically designed for the quantitative measurement of N-terminal pro-brain natriuretic peptide (NT-proBNP) levels in mouse serum, plasma, and tissue lysates. NT-proBNP is a valuable biomarker for various cardiac conditions, particularly heart failure, in mice. This kit allows researchers to accurately assess NT-proBNP levels, aiding in the diagnosis, prognosis, and monitoring of cardiac diseases in mouse models.
Q: What types of samples can be used with the Mouse NT-proBNP ELISA Kit?
The Mouse NT-proBNP ELISA Kit is validated for use with mouse serum, plasma, and tissue lysate samples. However, it is recommended to review the kit's instructions and protocols for specific requirements regarding sample collection, preparation, and dilution.
Q: Can the Mouse NT-proBNP ELISA Kit be used for other species besides mice?
The Mouse NT-proBNP ELISA Kit is specifically designed and optimized for the measurement of NT-proBNP in mouse samples. While it may cross-react with related proteins in other species, it is recommended to consult the kit manual or contact technical support to determine if it is suitable for use with samples from other species.
Q: Where can I find additional technical support or assistance with the Mouse NT-proBNP ELISA kit?
For any technical inquiries or assistance regarding the Mouse NT-proBNP ELISA kit, you can reach out to our team. They will be available to answer your questions and provide the necessary guidance to ensure a successful experiment.
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Mouse BNP / Brain Natriuretic Peptide ELISA Kit | |
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ELISA TYPE: | Competitive ELISA, Coated with Antigen |
SENSITIVITY: | 9.375pg/ml |
RANGE: | 15.625-1000pg/ml |