Defensins are cationic, anti-microbial peptides, produced by many cell types (e.g., leukocytes, epithelium, dendrites), and which play prominent roles in the innate immune response of mammals. Two classes described in humans, the alpha- and beta-defensins, range from 3.5 to 4.5kD and are stabilized by three intramolecular disulfide bonds differing by the ordering of the disulfide bonds in the mature peptides. To date, six alpha-defensins (also referred to as Human Neutrophil Peptides, HNP1-6) and six beta-defensins (HBD1-6) have been investigated. All have demonstrated broad-spectrum in vitro antimicrobial activity against bacteria, fungi and enveloped viruses – activities that are expected to be significant for in vivo protection against pathogens. Researchers have also investigated the ability of defensins to induce the release from tissues of cytokines and chemokines involved in inflammatory responses, thereby providing a regulatory role that bridges innate and adaptive immunity. Investigative research has led to the measurement of defensins in serum, saliva, milk, amniotic fluid and lung and cervicovaginal lavage, and culture media of cells and tissue of blood, lung, skin, bowel, muscle, cartilage and kidney, among other sample types.
Sample Type:Serum, saliva, milk, amniotic fluid and lung and cervicovaginal lavage, and culture media of cells and tissue of blood, lung, skin, bowel, muscle, cartilage and kidney, among other sample types.
Intended Use:The Human Beta Defensin 1 (BD-1) ELISA Kit is an in vitro immunoassay for research use in the quantification of BD-1 in cultures of human cells and in appropriately qualified samples from serum, saliva or other tissue fluids.
Range:Human saliva: 9.4-23.4ng/ml
Culture Medium: BD-1 was spiked into Sample Diluent with 10% Neonatal Bovine Serum at 4 levels, 100-800pg/ml. The mean recovery ranged from 105-118%, demonstrating linear dilution and equivalent quantification across the standard range.
Test Principle:The Human Beta Defensin 1 ELISA kit is based on the binding of Human Beta Defensin 1 in samples to two antibodies, one immobilized on the microtiter wells, and the other conjugated to biotin, which then binds to a streptavidin horseradish peroxidase (HRP) conjugate. After a washing step, chromogenic substrate is added and color is developed by the enzymatic reaction of HRP on the TMB substrate, which is directly proportional to the amount of BD-1 present in the sample. Stopping Solution is added to terminate the reaction, and absorbance at 450nm is then measured using an ELISA microtiter well reader. The concentration of BD-1 in samples is calculated from a standard curve of purified recombinant human BD-1 of designated concentration.
Kit Components:B0900-31A: Microtiter Strips 1x96 wells coated with Human BD1 ABB0900-31B: Standard 800pg/ml 3x1vialB0900-31C: Ab (Biotin), 100X 1x150ulB0900-31D: Streptavidin (HRP), 100X 1x150ulB0900-31E: Sample Diluent, 20X 1x10mlB0900-31F: Wash Solution, 100X 1x10mlB0900-31G: TMB Substrate 1x12mlB0900-31H: Stop Solution (1% H2SO4) 1x12ml
Storage and Stability:Store components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
