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ELK Biotechnology

ELK9769

CAF(C-terminal fragement of agrin) ELISA Kit

CAF(C-terminal fragement of agrin) ELISA Kit

Sensitivity

CAF(C-terminal fragement of agrin) ELISA Kit

Overview

Product name: Human CAF(C-terminal fragement of agrin) ELISA Kit
Reactivity: Human
Alternative Names: CAF; c-terminal fragement of agrin
Assay Type: Sandwich
Sensitivity: 13.1 pg/mL
Standard: 2000 pg/mL
Detection Range: 31.25-2000 pg/mL
Sample Type: Serum, plasma, tissue homogenates and other biological fluids
Assay Length: 3.5h
Research Area: Neuro science;Developmental science;Urology;
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 CAF. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human CAF. 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 CAF, 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 CAF 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
2000.00 2.255 2.173
1000.00 1.612 1.530
500.00 1.115 1.033
250.00 0.828 0.746
125.00 0.532 0.450
62.50 0.369 0.287
31.25 0.216 0.134
0.00 0.082 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 CAF and the recovery rates were calculated by comparing the measured value to the expected amount of CAF in samples.
Matrix Recovery range Average
serum(n=5) 79-97% 88%
EDTA plasma(n=5) 90-105% 97%
Heparin plasma(n=5) 87-95% 91%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of CAF 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) 94-106% 83-97% 88-96% 92-104%
EDTA plasma(n=5) 87-93% 95-106% 96-105% 95-103%
Heparin plasma(n=5) 93-101% 82-94% 86-96% 81-97%
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