Rat Phosphofructokinase, Liver (PFKL) ELISA Kit
Principle of the Assay
The microtiter plate provided in this kit has been pre-coated with an antibody specific to PFKL. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to PFKL. 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 PFKL, 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 PFKL 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 D-fructose 6-phosphate to fructose 1,6-bisphosphate by ATP, the first committing step of glycolysis (By similarity). Negatively regulates the phagocyte oxidative burst in response to bacterial infection by controlling cellular NADPH biosynthesis and NADPH oxidase-derived reactive oxygen species. Upon macrophage activation, drives the metabolic switch toward glycolysis, thus preventing glucose turnover that produces NADPH via pentose phosphate pathway (By similarity).
GENE ID | 25741 |
SWISS PROT | P30835 |
SYNONYMS |
PFK-B; 6-phosphofructokinase type B; Phosphofructo-1-kinase isozyme B; Phosphohexokinase |
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 PFKL were tested 20 times on one plate, respectively. |
SENSITIVITY | The minimum detectable dose was 0.58ng/mL. |
ASSAY RANGE | 1.56-100ng/mL |
SPECIFICITY | This assay has high sensitivity and excellent specificity for the detection of PFKL. No significant cross-reactivity or interference between PFKL and analogs was observed. Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between PFKL and all analogs, therefore, cross reactivity may still exist. |