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ELK Biotechnology

ELK9591

Human mTOR(MammalianTarget of Rapamycin) ELISA Kit

Human mTOR(MammalianTarget of Rapamycin) ELISA Kit

Sensitivity

Human mTOR(MammalianTarget of Rapamycin) ELISA Kit

Overview

Product name: Human mTOR(MammalianTarget of Rapamycin) ELISA Kit
Reactivity: Human
Alternative Names: FRAP1; RAFT1; RAPT1; Mechanistic target of rapamycin; FK506 binding protein 12-rapamycin associated protein 1
Assay Type: Sandwich
Sensitivity: 0.10 ng/mL
Standard: 10 ng/mL
Detection Range: 0.16-10 ng/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Cell proliferation
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 Human mTOR. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human mTOR. 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 Human mTOR, 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 Human mTOR 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
10.00 2.172 2.090
5.00 1.647 1.565
2.50 1.135 1.053
1.25 0.975 0.893
0.63 0.543 0.461
0.32 0.321 0.239
0.16 0.248 0.166
0.00 0.082 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 mTOR and the recovery rates were calculated by comparing the measured value to the expected amount of mTOR in samples.
Matrix Recovery range Average
serum(n=5) 80-95% 87%
EDTA plasma(n=5) 83-99% 95%
Heparin plasma(n=5) 94-107% 100%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of mTOR 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) 83-97% 95-104% 82-93% 81-96%
EDTA plasma(n=5) 78-92% 87-98% 89-96% 90-99%
Heparin plasma(n=5) 95-106% 87-101% 88-102% 89-103%
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