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Rat p-AMPK(Phosphorylated Adenosine Monophosphate Activated Protein Kinase) ELISA Kit
ELK0962
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48 T $320.00
96 T $458.00
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Overview

Product name: Rat p-AMPK(Phosphorylated Adenosine Monophosphate Activated Protein Kinase) ELISA Kit
Reactivity: Rat
Alternative Names: p-PRKAA1; p-AMPK; p-AMPKa1; p-AMPK alpha 1
Assay Type: Sandwich
Sensitivity: 1.24 ng/mL
Standard: 200 ng/mL
Detection Range: 3.13-200 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 Rat p-AMPK. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat 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 Rat 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 Rat 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
200.00 2.266 2.172
100.00 1.686 1.592
50.00 1.223 1.129
25.00 0.828 0.734
12.50 0.567 0.473
6.25 0.336 0.242
3.13 0.256 0.162
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 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) 83-98% 90%
EDTA plasma(n=5) 90-105% 97%
Heparin plasma(n=5) 87-95% 91%

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) 79-92% 87-103% 89-98% 80-101%
EDTA plasma(n=5) 89-96% 90-99% 85-96% 82-93%
Heparin plasma(n=5) 91-105% 87-98% 99-102% 89-102%