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Rat MGMT(O-6-Methylguanine DNA Methyltransferase) ELISA Kit
ELK7160
Size： Price：
48 T $320.00
96 T $458.00
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Overview

Product name: Rat MGMT(O-6-Methylguanine DNA Methyltransferase) ELISA Kit
Reactivity: Rat
Alternative Names: Methylated-DNA--Protein-Cysteine Methyltransferase; 6-O-methylguanine-DNA methyltransferase; Methylated-DNA--protein-cysteine methyltransferase
Assay Type: Sandwich
Sensitivity: 27 pg/mL
Standard: 5000 pg/mL
Detection Range: 78.13-5000 pg/mL
Sample Type: Tissue homogenates and other biological fluids.
Assay Length: 3.5h
Research Area: Signal transduction;Enzyme & Kinase;Tumor immunity;
Uniprot ID: P24528
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 MGMT. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat MGMT. 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 MGMT, 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 MGMT 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
5000.00 2.163 2.095
2500.00 1.484 1.416
1250.00 0.885 0.817
625.00 0.665 0.597
312.50 0.493 0.425
156.25 0.312 0.244
78.13 0.206 0.138
0.00 0.068 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 MGMT and the recovery rates were calculated by comparing the measured value to the expected amount of MGMT in samples.
Matrix Recovery range Average
serum(n=5) 90-104% 97%
EDTA plasma(n=5) 80-97% 88%
Heparin plasma(n=5) 87-99% 93%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of MGMT 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% 87-100% 85-94% 83-96%
EDTA plasma(n=5) 85-100% 84-98% 95-101% 88-102%
Heparin plasma(n=5) 79-92% 85-96% 80-98% 95-102%