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Human pNF-H(Phosphorylated Neurofilament H) ELISA Kit
ELK9302
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48 T $320.00
96 T $458.00
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Overview

Product name: Human pNF-H(Phosphorylated Neurofilament H) ELISA Kit
Reactivity: Human
Alternative Names: TC2; Transcobalamin II
Assay Type: Sandwich
Sensitivity: 29 pg/mL
Standard: 5000 pg/mL
Detection Range: 78.13-5000 pg/mL
Sample Type: Serum, plasma, tissue homogenates and other biological fluids.
Assay Length: 3.5h
Research Area: Neuro science
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 pNF-H. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human pNF-H. 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 pNF-H, 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 pNF-H in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (pg/mL) OD Corrected OD
5000.00 2.096 2.001
2500.00 1.587 1.492
1250.00 1.135 1.040
625.00 0.864 0.769
312.50 0.513 0.418
156.25 0.361 0.266
78.13 0.203 0.108
0.00 0.095 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 pNF-H and the recovery rates were calculated by comparing the measured value to the expected amount of pNF-H in samples.
Matrix Recovery range Average
serum(n=5) 82-97% 89%
EDTA plasma(n=5) 93-107% 100%
Heparin plasma(n=5) 95-107% 101%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of pNF-H 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) 89-96% 90-98% 87-98% 86-102%
EDTA plasma(n=5) 87-99% 99-105% 88-97% 93-103%
Heparin plasma(n=5) 81-98% 78-90% 91-99% 97-103%