Human Defensin Alpha 5, Paneth Cell Specific (DEFa5) ELISA Kit
Principle of the Assay
The microtiter plate provided in this kit has been pre-coated with an antibody specific to DEFa5. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to DEFa5. 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 DEFa5, 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 DEFa5 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
Host-defense peptide that maintains sterility in the urogenital system (PubMed:12021776, PubMed:12660734, PubMed:15616305, PubMed:22359618, PubMed:25354318, PubMed:30808760, PubMed:19589339, PubMed:22573326, PubMed:25782105). Has antimicrobial activity against a wide range of bacteria, including Gram-negative E.coli, P.aeruginosa and S.typhimurium, and Gram-positive E.aerogenes, S.aureus, B. cereus, E.faecium and L.monocytogenes (PubMed:12021776, PubMed:15616305, PubMed:22359618, PubMed:25354318, PubMed:30808760, PubMed:19589339, PubMed:22573326). Confers resistance to intestinal infection by S.typhimurium (PubMed:12660734). Exhibits antimicrobial activity against enteric commensal bacteria such as B.adolescentis, L.acidophilus, B.breve, L.fermentum, B.longum and S.thermophilus (PubMed:25354318). Binds to bacterial membranes and causes membrane disintegration (PubMed:25782105). Induces the secretion of the chemokine IL-8 by intestinal epithelial cells (PubMed:19589339). Binds to B.antracis lef/lethal factor, a major virulence factor from B.anthracis, and neutralizes its enzymatic activity (PubMed:22573326).
GENE ID | 1670 |
SWISS PROT | Q01523 |
SYNONYMS |
DEF-A5; DEF5; HD-5 |
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 DEFa5 were tested 20 times on one plate, respectively. |
SENSITIVITY | The minimum detectable dose was 1.31pg/mL. |
ASSAY RANGE | 3.12-200pg/mL |
SPECIFICITY | This assay has high sensitivity and excellent specificity for the detection of DEFa5. No significant cross-reactivity or interference between DEFa5 and analogs was observed. Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between DEFa5 and all analogs, therefore, cross reactivity may still exist. |