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Human 2,3-DPG(2,3-Diphosphoglycerate) ELISA Kit
ELK0778
Size： Price：
48 T $320.00
96 T $458.00
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Overview

Product name: Human 2,3-DPG(2,3-Diphosphoglycerate) ELISA Kit
Reactivity: Human
Alternative Names: 2,3DPG
Assay Type: Sandwich
Sensitivity: 0.14 µmol/mL
Standard: 20 µmol/mL
Detection Range: 0.32-20 µmol/mL
Sample Type: Serum, plasma, tissue homogenates, cell lysates and other biological fluids
Assay Length: 3.5h
Research Area: Metabolic pathway
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 2,3-DPG. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human 2,3-DPG. 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 2,3-DPG, 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 2,3-DPG in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (µmol/mL) OD Corrected OD
20.00 2.154 2.059
10.00 1.587 1.492
5.00 1.184 1.089
2.50 0.866 0.771
1.25 0.519 0.424
0.63 0.336 0.241
0.32 0.190 0.095
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 2,3-DPG and the recovery rates were calculated by comparing the measured value to the expected amount of 2,3-DPG in samples.
Matrix Recovery range Average
serum(n=5) 90-105% 97%
EDTA plasma(n=5) 80-92% 86%
Heparin plasma(n=5) 88-102% 95%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 2,3-DPG 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) 85-97% 90-99% 82-101% 93-105%
EDTA plasma(n=5) 85-97% 87-96% 82-95% 92-105%
Heparin plasma(n=5) 87-96% 92-105% 89-102% 92-101%