1 1

ELK Biotechnology

ELK4166

Human RFK(Riboflavin Kinase) ELISA Kit

Human RFK(Riboflavin Kinase) ELISA Kit

Size
Sensitivity
Detection range

Human RFK(Riboflavin Kinase) ELISA Kit

Overview

Product name: Human RFK(Riboflavin Kinase) ELISA Kit
Reactivity: Human
Alternative Names: RIFK; Flavokinase; ATP:riboflavin 5'-phosphotransferase
Assay Type: Sandwich
Sensitivity: 0.27 ng/mL
Standard: 50 ng/mL
Range: 0.79-50 ng/mL
Sample Type: Tissue homogenates, cell lysates 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 Human RFK. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human RFK. 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 RFK, 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 RFK 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
50.00 2.079 1.990
25.00 1.647 1.558
12.50 1.088 0.999
6.25 0.813 0.724
3.13 0.591 0.502
1.57 0.379 0.290
0.79 0.258 0.169
0.00 0.089 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 RFK and the recovery rates were calculated by comparing the measured value to the expected amount of RFK in samples.
Matrix Recovery range Average
serum(n=5) 87-93% 90%
EDTA plasma(n=5) 80-90% 85%
Heparin plasma(n=5) 84-106% 100%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of RFK 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) 88-102% 86-92% 81-93% 89-104%
EDTA plasma(n=5) 95-104% 79-92% 82-95% 86-97%
Heparin plasma(n=5) 85-92% 89-103% 97-105% 91-99%
전체 세부 정보 보기