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Human C4a(Complement Component 4a) ELISA Kit
ELK4951
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48 T $320.00
96 T $458.00
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Overview

Product name: Human C4a(Complement Component 4a) ELISA Kit
Reactivity: Human
Alternative Names: C4a anaphylatoxin
Assay Type: Sandwich
Sensitivity: 0.151 ng/mL
Standard: 20 ng/mL
Detection Range: 0.32-20 ng/mL
Sample Type: serum, plasma and other biological fluids
Assay Length: 3.5h
Research Area: Immune molecule;Endocrinology;
Uniprot ID: P0C0L4
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 C4a. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human C4a. 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 C4a, 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 C4a 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
20.00 2.266 2.177
10.00 1.695 1.606
5.00 1.135 1.046
2.50 0.863 0.774
1.25 0.529 0.440
0.63 0.262 0.173
0.32 0.196 0.107
0.00 0.089 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 C4a and the recovery rates were calculated by comparing the measured value to the expected amount of C4a in samples.
Matrix Recovery range Average
serum(n=5) 81-95% 88%
EDTA plasma(n=5) 85-97% 91%
Heparin plasma(n=5) 96-109% 102%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of C4a 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) 80-97% 81-92% 87-103% 82-93%
EDTA plasma(n=5) 95-104% 82-96% 92-105% 83-92%
Heparin plasma(n=5) 91-98% 83-101% 87-98% 95-102%