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Human pNF-H (Phosphorylated Neurofilament H) ELISA Kit
ELK4734
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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: NFH; 200 kDa neurofilament protein; Neurofilament triplet H protein; NEFH; Neurofilament, Heavy Polypeptide
Assay Type: Sandwich
Sensitivity: 5.9 pg/mL
Standard: 1000 pg/mL
Detection Range: 15.63-1000 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
1000.00 2.058 1.973
500.00 1.636 1.551
250.00 1.146 1.061
125.00 0.872 0.787
62.50 0.518 0.433
31.25 0.347 0.262
15.63 0.185 0.100
0.00 0.085 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) 85-97% 91%
EDTA plasma(n=5) 92-105% 98%
Heparin plasma(n=5) 82-95% 88%

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) 86-100% 93-108% 80-101% 87-96%
EDTA plasma(n=5) 96-105% 95-103% 95-103% 87-98%
Heparin plasma(n=5) 85-92% 79-96% 89-100% 85-96%