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ELK Biotechnology

ELK9598

Human C3d(ComplementFragment 3d) ELISA Kit

Human C3d(ComplementFragment 3d) ELISA Kit

Sensitivity

Human C3d(ComplementFragment 3d) ELISA Kit

Overview

Product name: Human C3d(ComplementFragment 3d) ELISA Kit
Reactivity: Human
Alternative Names: C3d
Assay Type: Sandwich
Sensitivity: 4.69 ng/mL
Standard: 500 ng/mL
Detection Range: 7.81-500 ng/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Immunology
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 C3d. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human C3d. 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 C3d, 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 C3d 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
500.00 1.946 1.850
250.00 1.663 1.567
125.00 1.185 1.089
62.50 0.876 0.780
31.25 0.543 0.447
15.63 0.357 0.261
7.82 0.215 0.119
0.00 0.096 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 C3d and the recovery rates were calculated by comparing the measured value to the expected amount of C3d in samples.
Matrix Recovery range Average
serum(n=5) 87-102% 94%
EDTA plasma(n=5) 95-107% 101%
Heparin plasma(n=5) 80-96% 88%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of C3d 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) 83-96% 93-107% 85-94% 87-101%
EDTA plasma(n=5) 99-106% 78-92% 87-95% 82-94%
Heparin plasma(n=5) 95-102% 87-103% 95-106% 87-101%
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