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Human Phospho AKT S473 (p-AKT) ELISA Kit
ELK0792
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48 T $320.00
96 T $458.00
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Overview

Product name: Human Phospho AKT S473 (p-AKT) ELISA Kit
Reactivity: Human
Alternative Names: p-AKT
Assay Type: Sandwich
Sensitivity: 52.5 pg/mL
Standard: 10000 pg/mL
Detection Range: 156.25-10000 pg/mL
Sample Type: tissue homogenates, cell lysates 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 Human p-AKT. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human p-AKT. 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 p-AKT, 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 p-AKT 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
10000.00 2.154 2.059
5000.00 1.638 1.543
2500.00 1.175 1.080
1250.00 0.871 0.776
625.00 0.577 0.482
312.50 0.315 0.220
156.25 0.258 0.163
0.00 0.095 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 p-AKT and the recovery rates were calculated by comparing the measured value to the expected amount of p-AKT in samples.
Matrix Recovery range Average
serum(n=5) 90-107% 98%
EDTA plasma(n=5) 82-98% 90%
Heparin plasma(n=5) 85-99% 92%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of p-AKT 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) 85-99% 79-93% 81-96% 85-96%
EDTA plasma(n=5) 87-105% 83-92% 82-96% 88-101%
Heparin plasma(n=5) 95-106% 85-104% 90-98% 89-105%