Alpha 1-Antitrypsin is a 52kD serine protease inhibitor (serpin) in blood, which protects tissue from enzymes from inflammatory cells, especially elastase. In certain acute phase inflammatory reactions, A1AT is elevated in order to limit the damage caused by activated neutrophil granulocytes and their enzyme elastase. Disorders of A1AT include A1AT deficiency, a hereditary disorder that can lead to severe tissue breakdown during inflammation. This may result in pulmonary emphysema and liver cirrhosis, in severe cases. Genetic variants of A1AT do occur.
Sample Type:Blood
Intended Use:The Alpha 1-Antitrypsin (A1AT) test kit is a highly sensitive two-site enzyme linked immunoassay (ELISA) for measuring A1AT in biological fluid of humans.
Test Principle:The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the Alpha 1-Antitrypsin present in samples reacts with the anti-Alpha 1-Antitrypsin antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-A1AT antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound A1AT. Following another washing step, the enzyme bound to the immunosorbent is assayed by the addition of a chromogenic substrate, 3,3’,5,5’- tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of A1AT in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of A1AT in the test sample. The quantity of A1AT in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.
Kit Components:Diluent concentrate (Running Buffer)One bottle containing 1x50ml of a 5X concentrated diluent running buffer.
Wash solution concentrateOne bottle containing 1x50ml of a 20X concentrated wash solution.
Enzyme-antybody conjugate 100XOne vial containing 1x150ul of affinity purified anti-Human Alpha 1-Antitrypsin antibody conjugated with horseradish peroxidase in a stabilizing buffer.
Chromogen-substrate solution One vial containing 1x12ml of 3,3’,5,5’-tetramethybenzidine (TMB) and hydrogen peroxide in citric acid buffer at pH 3.3.
Anti-human A1M ELISA micro plateTwelve removable eight (8) well micro well strips in well holder frame. Each well is coated with affinity purified anti-Human A1M.
Human alpha 1-antitrypsin calibratorOne vial containing a lyophilized Human Alpha 1-Antitrypsin calibrator.
Storage and Stability:Store all components at 4°C. Stable for at least 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
