| Field | Specification |
|---|---|
| UniProt # | |
| Applications | |
| Sensitivity | |
| Detection range | |
| Detection method | |
| Assay time | |
| Storage | |
| Shipping | |
| Catalog no. (Mfr.) | |
| Main SKU |
Scientific Background
This ELISA kit applies to the in vitro quantitative determination of Human NPHN concentrations in serum, plasma and other biological fluids.
Assay Principle
This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human NPHN. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human NPHN and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human NPHN, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human NPHN. You can calculate the concentration of Human NPHN in the samples by comparing the OD of the samples to the standard curve.
Performance Specifications
| Sensitivity | 0.1 ng/mL |
|---|---|
| Detection Range | 0.16-10 ng/mL |
| Total Assay Time | 3 h 30 min |
| Compatible Sample Types | Serum, plasma and other biological fluids |
| Species Reactivity | Human |
| Detection Method | Sandwich |
| Precision (CV) | Both intra-CV and inter-CV are < 10%. |
| Recovery Rate | 80%-120% |
| Storage | 2-8℃,12 months |
✓ Research-Grade Validation
Specificity
This kit recognizes Human NPHN in samples. No significant cross-reactivity or interference between Human NPHN and analogues was observed
Safety & Regulatory
Handle reagents in accordance with institutional biosafety guidelines. Refer to the Safety Data Sheet (SDS) for complete hazard and handling information. Contains components that may require special disposal procedures per local regulations.
This kit is validated for use with Serum, plasma and other biological fluids. For unlisted matrices (e.g., tissue lysate, urine), perform a spike-and-recovery experiment to confirm assay performance before generating reportable data. Sample dilution in the kit's provided diluent is recommended to minimize matrix interference.
The minimum detectable concentration (sensitivity) of this kit is 0.1 ng/mL. Values below this threshold should be reported as below the limit of detection (<LOD) and should not be extrapolated from the standard curve.
The total assay time from sample addition to absorbance reading is approximately 3 h 30 min, including all incubation, wash, and substrate steps. Hands-on time is typically 1–2 hours; most steps involve passive plate incubation. Plan the assay as a single uninterrupted session for best results.
Standard components of this Sandwich ELISA Kit typically include: pre-coated microplate (96-well strip format), lyophilized or liquid recombinant NPHN standard, detection antibody, streptavidin-HRP conjugate, TMB substrate, stop solution, wash buffer concentrate, and sample/standard diluent. Refer to the kit insert or datasheet for the exact component list and storage requirements.
This kit uses colorimetric (TMB/HRP) detection and requires a standard microplate absorbance reader capable of measuring at 450 nm. A reference wavelength of 570 nm or 630 nm is recommended to reduce background. No specialized fluorescence or luminescence reader is needed. Ensure the instrument is calibrated and the plate is clean and free of condensation before reading.
Tissue samples may contain endogenous or exogenous proteases during processing, leading to degradation of extracted proteins. Therefore, it's necessary to add protease inhibitors during processing to ensure the integrity of target proteins. If customers can keep samples cold and handle them quickly during processing, omitting the protease inhibitor may not have a significant effect. After preparation, samples should be tested promptly or immediately aliquoted and frozen at -20°C or -80°C.
What is the range of enzyme activity of your IL-2 freeze-dried powder
What is the principle of adding stop solution to stop color reaction in ELISA experiment?
Can’t Find What You’re Looking For? We can help you source the best match or customize an ELISA solution for your study. Options may include alternative target synonyms, different species reactivity, sample type/matrix compatibility (serum/plasma/lysate/supernatant), assay format (sandwich/competitive), sensitivity/range, detection chemistry (colorimetric/fluorescent/chemiluminescent), plate format (pre-coated/uncoated, strips vs full plate), and bulk or custom packaging. Click Talk to a Scientist to submit a request form, email us at support@biohippo.com, or explore our Research Services for additional support. Our team will be in contact with you shortly.
Neutrophil-to-lymphocyte ratio is elevated in acute hantavirus infection and correlates with markers of disease severity (2024) JOURNAL OF MEDICAL VIROLOGY. 10.1002/jmv.29759
Glomerular filtration barrier dysfunction in a self-limiting, RNA virus-induced glomerulopathy resembles findings in idiopathic nephrotic syndromes (2020) Scientific Reports. 10.1038/s41598-020-76050-0
Urinary and Serum Amino Acids May Be Associated with Podocyte, Proximal Tubule, and Renal Endothelial Injury in Early Diabetic Kidney Disease in Type 2 Diabetes Mellitus Patients (2025) Biomedicines. 10.3390/biomedicines13030675
Proximal Tubule Dysfunction Is Associated with Podocyte Damage Biomarkers Nephrin and Vascular Endothelial Growth Factor in Type 2 Diabetes Mellitus Patients: A Cross-Sectional Study (2014) PLoS One. 10.1371/journal.pone.0112538
Glomerular Injury Is Associated with Severe Courses of Orthohantavirus Infection (2024) Pathogens. 10.3390/pathogens13080693
The Use of Kidney Biomarkers, Nephrin and KIM-1, for the Detection of Early Glomerular and Tubular Damage in Patients with Acromegaly: A Case–Control Pilot Study (2024) Diseases. 10.3390/diseases12090211
Podocyturia parallels proximal tubule dysfunction in type 2 diabetes mellitus patients independently of albuminuria and renal function decline: A cross-sectional study (2017) JOURNAL OF DIABETES AND ITS COMPLICATIONS. 10.1016/j.jdiacomp.2017.01.007
A pilot study of anti-nephrin antibodies in podocytopaties among adults (2023) NEPHROLOGY. 10.1111/nep.14249
Urinary podocyte-associated mRNA levels correlate with proximal tubule dysfunction in early diabetic nephropathy of type 2 diabetes mellitus (2017) Diabetology & Metabolic Syndrome. 10.1186/s13098-017-0228-y
Urinary Nephrin: A Potential Biomarker of Early Glomerular Injury in a Cohort of Pregnant Women Attending Routine Antenatal Care Services (2024) International Journal of Nephrology. 10.1155/2024/9089557
Therapy with atorvastatin versus rosuvastatin reduces urinary podocytes, podocyte-associated molecules, and proximal tubule dysfunction biomarkers in patients with type 2 diabetes mellitus: a pilot study (2016) RENAL FAILURE. 10.1080/0886022X.2016.1254657
