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ELK Biotechnology

ELK6916

Rat NEFH(Neurofilament, Heavy Polypeptide) ELISA Kit

Rat NEFH(Neurofilament, Heavy Polypeptide) ELISA Kit

$458.00 USD
$458.00 USD
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Size
Sensitivity
Detection range

Rat NEFH(Neurofilament, Heavy Polypeptide) ELISA Kit

Overview

Product name: Rat pNF-H (Phosphorylated Neurofilament H) ELISA Kit
Reactivity: Rat
Alternative Names: NFH; 200 kDa neurofilament protein; Neurofilament triplet H protein; NEFH; Neurofilament, Heavy Polypeptide
Assay Type: Sandwich
Sensitivity: 5.7 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 Rat pNF-H. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat 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 Rat 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 Rat pNF-H 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
1000.00 2.252 2.169
500.00 1.502 1.419
250.00 1.157 1.074
125.00 0.839 0.756
62.50 0.593 0.510
31.25 0.345 0.262
15.63 0.186 0.103
0.00 0.083 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) 95-108% 101%
EDTA plasma(n=5) 83-96% 89%
Heparin plasma(n=5) 88-102% 95%

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) 86-97% 88-95% 92-101% 87-98%
EDTA plasma(n=5) 91-99% 85-93% 82-95% 93-99%
Heparin plasma(n=5) 79-91% 87-96% 83-101% 82-95%
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