Human Riboflavin Kinase (RFK) ELISA Kit
Principle of the Assay
The microtiter plate provided in this kit has been pre-coated with an antibody specific to RFK. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to RFK. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After the TMB substrate solution is added, only those wells that contain 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 RFK in the samples is then determined by comparing the O.D. of the samples to the standard curve.
For Use with serum, plasma, and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Target Information
Catalyzes the phosphorylation of riboflavin (vitamin B2) to form flavin-mononucleotide (FMN), hence rate-limiting enzyme in the synthesis of FAD. Essential for TNF-induced reactive oxygen species (ROS) production. Through its interaction with both TNFRSF1A and CYBA, physically and functionally couples TNFRSF1A to NADPH oxidase. TNF-activation of RFK may enhance the incorporation of FAD in NADPH oxidase, a critical step for the assembly and activation of NADPH oxidase.
GENE ID | 55312 |
SWISS PROT | Q969G6 |
SYNONYMS |
RIFK; Flavokinase; ATP:riboflavin 5'-phosphotransferase |
Materials Supplied
Kit Components | 96 Wells Quantity/Size |
---|---|
Pre-coated, ready-to-use 96-well strip plate | 1 plate |
Plate sealer for 96 wells | 2 |
Standard |
2 tubes |
Diluent buffer | 1 bottle |
Detection Reagent A | 1 bottle |
Detection Reagent B | 1 bottle |
TMB Substrate | 1 tube |
Stop Solution | 1 tube |
Wash Buffer (30 ℅ concentrate) | 1 tube |
Product data sheet | 1 copy |
Storage
Storage | The TMB Substrate, Wash Buffer (30X concentrate), and the Stop Solution should be stored at 4°C upon receipt, while the other items should be stored at -20°C. |
Performance Characteristics
REPEATABILITY |
Intra-assay Precision (Precision within an assay): 3 samples with low, middle, and high-level RFK were tested 20 times on one plate, respectively. |
SENSITIVITY | The minimum detectable dose was 0.27ng/mL. |
ASSAY RANGE | 0.781-50ng/mL |
SPECIFICITY | This assay has high sensitivity and excellent specificity for the detection of RFK. No significant cross-reactivity or interference between RFK and analogs was observed. Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between RFK and all analogs, therefore, cross reactivity may still exist. |