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ELK Biotechnology

ELK9525

Mouse BCKDHB(2-oxoisovalerate dehydrogenase subunit beta, mitochondrial) ELISA Kit

Mouse BCKDHB(2-oxoisovalerate dehydrogenase subunit beta, mitochondrial) ELISA Kit

Sensitivity

Mouse BCKDHB(2-oxoisovalerate dehydrogenase subunit beta, mitochondrial) ELISA Kit

Overview

Product name: Mouse BCKDHB(2-oxoisovalerate dehydrogenase subunit beta, mitochondrial) ELISA Kit
Reactivity: Mouse
Alternative Names: 2-oxoisovalerate dehydrogenase subunit beta, mitochondrial; Branched-chain alpha-keto acid dehydrogenase E1 component beta chain; BCKDE1B; BCKDH E1-beta; Bckdhb
Assay Type: Sandwich
Sensitivity: 12.2 pg/mL
Standard: 2000 pg/mL
Detection Range: 31.25-2000 pg/mL
Sample Type: Serum, plasma, tissue homogenates and other biological fluids
Assay Length: 3.5h
Research Area: Enzyme & Kinase;
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 BCKDHB. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Mouse BCKDHB. 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 BCKDHB, 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 BCKDHB 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
2000.00 2.096 1.998
1000.00 1.587 1.489
500.00 1.211 1.113
250.00 0.839 0.741
125.00 0.545 0.447
62.50 0.357 0.259
31.25 0.256 0.158
0.00 0.098 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 BCKDHB and the recovery rates were calculated by comparing the measured value to the expected amount of BCKDHB in samples.
Matrix Recovery range Average
serum(n=5) 80-93% 86%
EDTA plasma(n=5) 86-99% 92%
Heparin plasma(n=5) 92-107% 99%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of BCKDHB 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) 87-98% 85-96% 82-97% 88-105%
EDTA plasma(n=5) 85-92% 90-96% 79-102% 81-99%
Heparin plasma(n=5) 94-106% 83-97% 79-96% 85-99%
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