Human NPHN(Nephrin) ELISA Kit (E-EL-H1901)

Name: Human NPHN(Nephrin) ELISA Kit
Brand: Elabscience
SKU: E-EL-H1901
Price: 150.0 USD
Availability: InStock

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Size	Price	Qty
96T	$ 609.00	- +
48T	$ 487.00	- +
24T	$ 150.00	- +
96T*5	Inquire	/
96T*10	Inquire	/

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Product Summary

Sensitivity	0.1 ng/mL
Detection Range	0.16-10 ng/mL
Sample Volume	100 μL
Total Assay Time	3 h 30 min
Reactivity	Human
Specificity	This kit recognizes Human NPHN in samples. No significant cross-reactivity or interference between Human NPHN and analogues was observed
Recovery	80%-120%
Sample Type	Serum, EDTA plasma, heparinized plasma, breast milk, Urine and Cell culture supernatant
Detection Method	Colorimetric method, ELISA, Sandwich
Assay Type	Sandwich-ELISA
Size	96T / 48T / 24T / 96T5 / 96T10
Storage	2-8℃
Expiration Date	12 months

Performance

Typical data

The following data was generated by the Quality Control Department, under controlled laboratory conditions (ambient temperature: 18-25 °C, relative humidity: 35-75%) using standardized procedures (TMB reaction at 37 °C in the dark for 15 minutes, followed by termination and OD measurement). These values are provided for reference only. Actual results may vary due to differences in laboratory conditions, operator technique, and equipment. Users are required to generate a standard curve using their own experimental data.

ng/mL	OD1	OD2	Mean OD	Corrected OD
10	2.328	2.288	2.308	2.224
5	1.577	1.581	1.579	1.495
2.5	0.935	0.945	0.940	0.856
1.25	0.525	0.499	0.512	0.428
0.63	0.317	0.337	0.327	0.243
0.32	0.192	0.186	0.189	0.105
0.16	0.145	0.135	0.140	0.056
0	0.087	0.081	0.084	-

Precision

Intra-assay Precision (Within-run Precision): Three samples representing low, mid, and high concentrations of Human NPHN were tested 20 times on a single plate.

Inter-assay Precision (Between-run Precision): Three samples representing low, mid, and high concentrations of Human NPHN were tested on three separate plates, with 20 replicates per plate, to assess variability among assays.

/	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
Numbers	20	20	20	20	20	20
Mean（ng/mL）	0.550	1.500	4.550	0.550	1.500	4.630
Standard deviation	0.030	0.080	0.220	0.040	0.120	0.330
CV（%）	5.350	5.140	4.820	7.050	8.320	7.190

Recovery

The recovery of Human NPHN was evaluated by spiking samples at low, mid, and high concentrations across the assay range in various sample matrices.
The assay performance was assessed by comparing the measured concentrations to the expected spiked amounts to determine the percent recovery.

Sample Type	Range （%）	Average Recovery（%）
Cell culture media （n=8）	89-102	95
Serum （n=8）	96-108	101
EDTA plasma （n=8）	90-106	96

Linearity

Linearity of the assay was evaluated by spiking samples with high concentrations of Human NPHN and performing serial dilutions using Standard & Sample Diluent to produce concentrations spanning the assay's dynamic range. The measured values were then compared to the expected concentrations to assess the linearity of response.

/	/	Cell culture media （n=5）	EDTA plasma （n=5）	Serum （n=5）
1:2	Range	94-107	92-107	88-98
1:2	Average	99	99	93
1:4	Range	83-98	83-94	87-100
1:4	Average	90	88	94
1:8	Range	83-97	83-97	88-101
1:8	Average	89	90	95

Stability

Each kit batch is subjected to accelerated stability testing and real-time stability monitoring. Sample performance is evaluated after storage at 37 °C for 10 days to assess the impact of elevated temperature on assay reliability and reagent integrity.

/	Variation range of 37°C mean concentration / 2-8°C mean
Sample 1（n=16）	88.78-104.88
Sample 2（n=16）	92.89-103.54

Test 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.

Background

Ghrelin is a 28-amino-acid peptide hormone that is the endogenous ligand of the growth hormone secretagogue receptor (GHSR). It is synthesized predominantly by the X/A-like endocrine cells of the gastric mucosa from a prepropeptide encoded by the GHRL gene, circulates in acylated and des-acylated forms, and is released in a manner regulated by nutritional state and feeding behavior, rising before meals and falling after food intake. Acylation of the serine at position 3 with an octanoic acid moiety is essential for receptor binding and biological activity and is catalyzed by ghrelin O-acyltransferase. Acting through GHSR in the pituitary and hypothalamus, ghrelin stimulates growth hormone release and is a potent orexigenic signal that increases food intake, reduces fat utilization, promotes adiposity and regulates energy balance in opposition to leptin, while also stimulating gastric motility, acid secretion and reward-related feeding circuits. Altered ghrelin signaling is implicated in obesity, cachexia, anorexia nervosa, Prader-Willi syndrome and postoperative or chemotherapy-induced appetite loss, and total and acylated ghrelin differ in their physiological meaning. Measurement of the hormone is widely used in studies of appetite regulation, energy homeostasis and growth hormone physiology, and detection of the acylated form in particular supports metabolic, endocrine and gastrointestinal research and evaluation of interventions affecting feeding behavior and body composition.