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Rat IL-12p40(Interleukin 12 p40) Microsample ELISA Kit
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ELK2509MS
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48 T $320.00
96 T $458.00
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Overview

Product name: Rat IL-12p40(Interleukin 12 p40) Microsample ELISA Kit
Reactivity: Rat
Alternative Names: IL-12B; p40; IL12B; CLMF2; NKSF2; Natural Killer Cell Stimulatory Factor 2; Cytotoxic Lymphocyte Maturation Factor 2; Interleukin 23, P40 Subunit
Assay Type: Sandwich
Sensitivity: 5.8 pg/mL
Standard: 1000 pg/mL
Detection Range: 15.63-1000 pg/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Cytokine;Infection immunity;Immune molecule;
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 Rat IL-12p40. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat IL-12p40. 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 Rat IL-12p40, 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 Rat IL-12p40 in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (pg/mL) OD Corrected OD
1000.00 2.061 1.967
500.00 1.527 1.433
250.00 1.120 1.026
125.00 0.762 0.668
62.50 0.561 0.467
31.25 0.340 0.246
15.63 0.170 0.076
0.00 0.094 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 IL-12p40 and the recovery rates were calculated by comparing the measured value to the expected amount of IL-12p40 in samples.
Matrix Recovery range Average
serum(n=5) 95-110% 103%
EDTA plasma(n=5) 86-128% 107%
Heparin plasma(n=5) 88-99% 94%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of IL-12p40 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) 82-114% 81-120% 86-119% 86-108%
EDTA plasma(n=5) 89-102% 93-107% 90-118% 93-111%
Heparin plasma(n=5) 91-97% 80-115% 84-109% 95-100%

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