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ELK Biotechnology

ELK9502

Human NEDD4-L(E3 ubiquitin-protein ligase NEDD4-like) ELISA Kit

Human NEDD4-L(E3 ubiquitin-protein ligase NEDD4-like) ELISA Kit

Sensitivity

Human NEDD4-L(E3 ubiquitin-protein ligase NEDD4-like) ELISA Kit

Overview

Product name: Human NEDD4-L(E3 ubiquitin-protein ligase NEDD4-like) ELISA Kit
Reactivity: Human
Alternative Names: NED4L;NEDD4 2
Assay Type: Sandwich
Sensitivity: 0.119 ng/mL
Standard: 20 ng/mL
Detection Range: 0.32-20 ng/mL
Sample Type: serum, plasma, tissue homogenates and other biological fluids
Assay Length: 3.5h
Research Area: Enzyme & Kinase;
Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Human NEDD4-L. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human NEDD4-L. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Human NEDD4-L, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Human NEDD4-L in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (ng/mL) OD Corrected OD
20.00 2.266 2.179
10.00 1.675 1.588
5.00 1.169 1.082
2.50 0.896 0.809
1.25 0.559 0.472
0.63 0.357 0.270
0.32 0.175 0.088
0.00 0.087 0.000

Precision

Intra-assay Precision (Precision within an assay)：CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays)：CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery

Matrices listed below were spiked with certain level of recombinant NEDD4-L and the recovery rates were calculated by comparing the measured value to the expected amount of NEDD4-L in samples.
Matrix Recovery range Average
serum(n=5) 82-95% 88%
EDTA plasma(n=5) 82-95% 88%
Heparin plasma(n=5) 80-95% 87%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of NEDD4-L and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 82-96% 85-97% 93-101% 85-98%
EDTA plasma(n=5) 90-101% 92-103% 81-99% 94-106%
Heparin plasma(n=5) 83-97% 93-103% 87-98% 85-96%
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