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Mouse Anti-KLH IgM(Keyhole Limpet Hemocyanin IgM) ELISA Kit
ELK0110
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48 T $320.00
96 T $458.00
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Overview

Product name: Mouse Anti-KLH IgM(Keyhole Limpet Hemocyanin IgM) ELISA Kit
Reactivity: Mouse
Alternative Names: Keyhole limpet IgM
Assay Type: Sandwich
Sensitivity: 0.51 ng/mL
Standard: 100 ng/mL
Detection Range: 1.57-100 ng/mL
Sample Type: Serum, plasma, tissue homogenates
Assay Length: 3.5h
Research Area: Immunology
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 Anti-KLH IgM. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Mouse Anti-KLH IgM. 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 Anti-KLH IgM, 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 Anti-KLH IgM 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 2.379 2.298
50.00 1.517 1.436
25.00 1.151 1.070
12.50 0.618 0.537
6.25 0.508 0.427
3.13 0.342 0.261
1.57 0.234 0.153
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 Anti-KLH IgM and the recovery rates were calculated by comparing the measured value to the expected amount of Anti-KLH IgM in samples.
Matrix Recovery range Average
serum(n=5) 85-97% 91%
EDTA plasma(n=5) 95-107% 101%
Heparin plasma(n=5) 82-95% 88%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Anti-KLH IgM 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) 85-94% 87-101% 86-97% 88-95%
EDTA plasma(n=5) 88-97% 90-99% 85-97% 79-101%
Heparin plasma(n=5) 78-94% 97-105% 88-101% 82-96%