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Mouse p-AMPK(Phosphorylated Adenosine Monophosphate Activated Protein Kinase) ELISA Kit
ELK9450
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48 T $320.00
96 T $458.00
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Overview

Product name: Mouse p-AMPK(Phosphorylated Adenosine Monophosphate Activated Protein Kinase) ELISA Kit
Reactivity: Mouse
Alternative Names: p-PRKAA1; p-AMPK; p-AMPKa1; p-AMPK alpha 1
Assay Type: Sandwich
Sensitivity: 0.59 ng/mL
Standard: 100 ng/mL
Detection Range: 1.57-100 ng/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates 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 p-AMPK. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Mouse p-AMPK. 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 p-AMPK, 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 p-AMPK 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 2.096 2.010
50.00 1.575 1.489
25.00 1.211 1.125
12.50 0.826 0.740
6.25 0.556 0.470
3.13 0.389 0.303
1.57 0.236 0.150
0.00 0.086 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-AMPK and the recovery rates were calculated by comparing the measured value to the expected amount of p-AMPK in samples.
Matrix Recovery range Average
serum(n=5) 83-95% 89%
EDTA plasma(n=5) 89-103% 96%
Heparin plasma(n=5) 87-99% 93%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of p-AMPK 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) 87-101% 95-102% 78-97% 85-96%
EDTA plasma(n=5) 82-93% 86-95% 81-96% 79-91%
Heparin plasma(n=5) 90-99% 85-92% 88-96% 87-98%