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ELK Biotechnology

ELK7917

Pig C5b-9(Terminal Complement Complex C5b-9) ELISA Kit

Pig C5b-9(Terminal Complement Complex C5b-9) ELISA Kit

Size
Sensitivity
Detection range

Pig C5b-9(Terminal Complement Complex C5b-9) ELISA Kit

Overview

Product name: Pig sC5b-9(Soluble Terminal Complement Complex C5b-9) ELISA Kit
Reactivity: Pig
Alternative Names: MAC; Membrane Attack Complex; C5b-9; Terminal Complement Complex C5b-9; C5b9
Assay Type: Sandwich
Sensitivity: 0.32 ng/mL
Standard: 50 ng/mL
Range: 0.79-50 ng/mL
Sample Type: Plasma,  tissue homogenates and other biological fluids
Assay Length: 3.5h
Research Area: Signal transduction;Metabolic pathway;Apoptosis;Infection immunity;
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 Pig sC5b-9. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Pig 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 Pig 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 Pig 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
50.00 2.043 1.971
25.00 1.643 1.571
12.50 1.186 1.114
6.25 0.752 0.680
3.13 0.527 0.455
1.57 0.321 0.249
0.79 0.206 0.134
0.00 0.072 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) 95-107% 101%
EDTA plasma(n=5) 77-95% 86%
Heparin plasma(n=5) 84-97% 90%

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) 85-92% 88-96% 87-98% 95-102%
EDTA plasma(n=5) 85-101% 84-98% 85-93% 86-98%
Heparin plasma(n=5) 86-93% 91-105% 87-98% 89-94%
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