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ELK Biotechnology

ELK9699

Rabbit sC5b-9(Soluble Terminal Complement Complex C5b-9) ELISA Kit

Rabbit sC5b-9(Soluble Terminal Complement Complex C5b-9) ELISA Kit

Sensitivity

Rabbit sC5b-9(Soluble Terminal Complement Complex C5b-9) ELISA Kit

Overview

Product name: Rabbit sC5b-9(Soluble Terminal Complement Complex C5b-9) ELISA Kit
Reactivity: Rabbit
Alternative Names: MAC; Membrane Attack Complex; C5b-9; Terminal Complement Complex C5b-9; C5b9
Assay Type: Sandwich
Sensitivity: 0.10 ng/mL
Standard: 10 ng/mL
Detection Range: 0.16-10 ng/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Immunology；Metabolism；Signal Transduction
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 Rabbit sC5b-9. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rabbit sC5b-9. 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 Rabbit sC5b-9, 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 Rabbit sC5b-9 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
10.00 2.054 1.959
5.00 1.612 1.517
2.50 1.211 1.116
1.25 0.933 0.838
0.63 0.559 0.464
0.32 0.385 0.290
0.16 0.186 0.091
0.00 0.095 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 sC5b-9 and the recovery rates were calculated by comparing the measured value to the expected amount of sC5b-9 in samples.
Matrix Recovery range Average
serum(n=5) 85-101% 93%
EDTA plasma(n=5) 78-92% 85%
Heparin plasma(n=5) 87-99% 93%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of sC5b-9 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) 99-106% 78-92% 83-97% 85-93%
EDTA plasma(n=5) 98-105% 87-98% 91-99% 85-93%
Heparin plasma(n=5) 82-95% 93-99% 85-97% 83-96%
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