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ELK Biotechnology

ELK9632

Rat TRACP-5b(TartrateResistant Acid Phosphatase 5b) ELISA Kit

Rat TRACP-5b(TartrateResistant Acid Phosphatase 5b) ELISA Kit

Sensitivity

Rat TRACP-5b(TartrateResistant Acid Phosphatase 5b) ELISA Kit

Overview

Product name: Rat TRACP-5b(TartrateResistant Acid Phosphatase 5b) ELISA Kit
Reactivity: Rat
Alternative Names: ACP5;TRAP5b;SPENCDI
Assay Type: Sandwich
Sensitivity: 0.38 ng/mL
Standard: 40 ng/mL
Detection Range: 0.63-40 ng/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Immunology;Cardiovascular
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 TRACP-5b. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat TRACP-5b. 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 TRACP-5b, 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 TRACP-5b 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
40.00 2.035 1.941
20.00 1.593 1.499
10.00 1.128 1.034
5.00 0.826 0.732
2.50 0.614 0.520
1.25 0.352 0.258
0.63 0.194 0.100
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 TRACP-5b and the recovery rates were calculated by comparing the measured value to the expected amount of TRACP-5b in samples.
Matrix Recovery range Average
serum(n=5) 80-95% 87%
EDTA plasma(n=5) 85-97% 91%
Heparin plasma(n=5) 78-95% 86%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of TRACP-5b 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) 95-103% 87-103% 91-102% 85-92%
EDTA plasma(n=5) 89-96% 90-98% 87-98% 92-104%
Heparin plasma(n=5) 85-95% 82-91% 85-97% 89-102%
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