BAFF (SOLUBLE) ELISA KIT
INTENDED USE
The BAFF (Soluble) ELISA Kit is intended for the quantitative determination of soluble (human) BAFF in serum and cell culture supernatant. It is for in vitro diagnostic use only.
INTRODUCTION
BAFF (B cell activating factor belonging to the TNF family, also known as BLyS or TALL1) is a cytokine expressed predominantly by cells of the immune system such as neutrophils, monocytes, macrophages, dendritic cells, follicular dendritic cells, activated T cells and some malignant B cells.
BAFF binds three distinct receptors (BAFFR, TACI and BCMA) predominantly expressed on B cells, although activated T cells also express BAFFR. BAFF is a master regulator of peripheral B cell survival, and also acts in processes such as immunoglobulin isotype switch and B cell co-stimulation. Beside its major role in B cell biology, BAFF co-stimulates activated T cells. Deregulated expression of this membrane-bound protein, which can readily be released in a soluble form by proteolytic cleavage, leads to autoimmune disorders in mice. In the human, elevated levels of soluble BAFF have been detected in the serum of patients with various autoimmune diseases, such as rheumatoid arthritis (RA), Sjögren's syndrome (SS) and systemic lupus erythematosus (SLE). BAFF levels are also elevated in patients with multiple myeloma and B-cell chronic lymphoid leukemia (B-CCL).
PRINCIPLE OF THE TEST
BAFF (Soluble) ELISA Kit
BAFF (Soluble) ELISA Kit is intended for the quantitative determination of soluble (human) BAFF in serum and cell culture supernatant. This BAFF (Soluble) ELISA Kit assay is a sandwich (ELISA) for the direct measurement of human BAFF in serum and cell culture supernatants.
Standards, controls and samples containing human BAFF are added to wells of microplate coated with a high affinity monoclonal anti-human BAFF antibody. During the first incubation period, the antibody immobilized on the wall of the microtiter wells captures BAFF in the patient samples or in the cell culture supernatants. After washing away the unbound components from samples, a detection antibody (monoclonal anti-BAFF antibody) is added to each well. During the incubation step, the detection antibody is bound to the captured BAFF. A peroxidase-conjugated antibody is then added to each microtiter well and a “sandwich” of capture antibody – human BAFF – detection antibody - Peroxidase conjugate is formed. Tetramethylbenzidine (TMB) is used as a substrate for peroxidase. Finally, an acidic stop solution is added to terminate the reaction. The intensity of the yellow color is directly proportional to the BAFF concentration of sample. A dose response curve of absorbance unit (optical density, OD at 450 nm) vs. concentration is generated; using the values obtained from standard. BAFF present in the patient samples or cell culture supernatants, is determined directly from this curve.
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