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Mouse C5a(Complement Component 5a) Microsample ELISA Kit
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ELK1052MS
Size： Price：
48 T $265.00
96 T $376.00
96 T*5 $1598.00
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Overview

Product name: Mouse C5a(Complement Component 5a) Microsample ELISA Kit
Reactivity: Mouse
Alternative Names: Complement Component 5a
Assay Type: Sandwich
Sensitivity: 0.59 ng/mL
Standard: 100 ng/mL
Detection Range: 1.57-100 ng/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Infection immunity;Immune molecule;Autoimmunity;
Uniprot ID: P06684
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 Mouse C5a. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Mouse C5a. 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 Mouse C5a, 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 Mouse C5a 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
100.00 1.968 1.887
50.00 1.523 1.442
25.00 1.175 1.094
12.50 0.810 0.729
6.25 0.515 0.434
3.13 0.316 0.235
1.56 0.216 0.135
0.00 0.081 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 C5a and the recovery rates were calculated by comparing the measured value to the expected amount of C5a in samples.
Matrix Recovery range Average
serum(n=5) 88-94% 91%
EDTA plasma(n=5) 94-117% 106%
Heparin plasma(n=5) 94-109% 102%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of C5a 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) 82-119% 80-85% 92-100% 85-110%
EDTA plasma(n=5) 94-119% 85-102% 84-93% 80-114%
Heparin plasma(n=5) 85-99% 87-98% 90-113% 84-119%

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