The Human Beta Defensin 3 (BD-3) ELISA Kit is an in vitro immunoassay for research use in the quantification of BD-3 in cultures of human cells and in appropriately qualified samples from serum, saliva, or other tissue fluids. The kit is suitable for testing a variety of sample types, in accordance with appropriate validation of linearity and recovery.
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 have been described in humans, the alpha- and beta-defensins, which range from 3.5-4.5kD 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 and/or adaptive immunity, 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.
Principle of the Test:The assay is based on the binding of human Beta Defensin 3 in samples to two antibodies, one immobilized on the microtiter wells, and the other conjugated to biotin, which then binds to a streptavidin (HRP). 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-3 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-3 in samples is calculated from a standard curve of purified recombinant human BD-3 of designated concentration.
Range:12.5-400pg/ml
Kit Components:B0902-06A: Microwell Strips, 1x96 wells (12x8-well strips). Coated with purified anti-human BD3 antibodies. Return unused strips to the pouch with desiccant; re-seal and store refrigerated
B0902-06B: Standard, 400pg/ml, 2x1 vial. Recombinant human BD3. Supplied as a lyophilized powder from buffer, protein stabilizers
B0902-06C: Ab (Biotin), 100X, 1x150ul. Biotinylated anti-human BD3. Supplied as a liquid in buffer, protein, detergents, 0.02% Bronidox. B0902-06D: Streptavidin (HRP), 100X, 1x150ul. Supplied as a liquid in buffer with protein, detergents, 0.02% Bronidox
B0902-06E: Sample Diluent, 10X, 1x10ml
B0902-06F: Biotin, HRP Diluent, 1x22ml. Supplied as a liquid in buffer, protein, anti-microbials
B0902-06G: Wash Solution, 100X, 1x10ml
B0902-06H: TMB Solution, 1x12ml. Chromogenic substrate for HRP containing TMB and peroxide
Storage and Stability:Store all components at 4°C. Stable for 6 months after receipt For maximum recovery of product, centrifuge the original vial prior to removing the cap.
