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ELK Biotechnology

ELK9457

Rat HK1(Hexokinase 1) ELISA Kit

Rat HK1(Hexokinase 1) ELISA Kit

Sensitivity

Rat HK1(Hexokinase 1) ELISA Kit

Overview

Product name: Rat HK1(Hexokinase 1) ELISA Kit
Reactivity: Rat
Alternative Names: HK1-Ta; HK1-Tb; HK1-Tc; HKI; HXK1; Brain form hexokinase
Assay Type: Sandwich
Sensitivity: 27.5 pg/mL
Standard: 4000 pg/mL
Detection Range: 62.5-4000 pg/mL
Sample Type: Serum, plasma, tissue homogenates, cell lysates and other biological fluids
Assay Length: 3.5h
Research Area: Enzyme & Kinase;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 Rat HK1. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat HK1. 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 HK1, 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 HK1 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
4000.00 1.929 1.841
2000.00 1.668 1.580
1000.00 1.135 1.047
500.00 0.827 0.739
250.00 0.521 0.433
125.00 0.321 0.233
62.50 0.213 0.125
0.00 0.088 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 HK1 and the recovery rates were calculated by comparing the measured value to the expected amount of HK1 in samples.
Matrix Recovery range Average
serum(n=5) 80-95% 87%
EDTA plasma(n=5) 80-96% 88%
Heparin plasma(n=5) 87-99% 93%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HK1 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% 82-96% 85-98% 95-104%
EDTA plasma(n=5) 93-102% 79-94% 87-98% 83-97%
Heparin plasma(n=5) 95-104% 95-106% 85-104% 90-98%
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