The Anatoxin-a ELISA is an immunoassay for the quantitative and sensitive detection of Anatoxin-a in water samples.
The Anatoxin-a ELISA is an immunoassay for the quantitative and sensitive detection of Anatoxin-a in water samples. The test is a direct competitive ELISA based on the recognition of Anatoxin-a by a monoclonal antibody. Anatoxin-a, when present in a sample, and an Anatoxin-a-enzyme conjugate compete for the binding sites of mouse anti-Anatoxin-a antibodies in solution. The Anatoxin-a antibodies are then bound by a second antibody (anti-mouse) immobilized on the microtiter plate. After a washing step and addition of the substrate solution, a color signal is generated. The intensity of the blue color is inversely proportional to the concentration of Anatoxin-a present in the sample. The color reaction is stopped after a specified time and the color is evaluated using an ELISA reader. The concentrations of the samples are determined by interpolation using the standard curve constructed with each run.
No. of tests
0.1 ppb (μg/L)
1. Microtiter plate coated with a secondary antibody (anti-mouse), in a re-sealable aluminum pouch 2. Lyophilized Anatoxin-a-HRP Enzyme Conjugate, 3 vials 3. Conjugate Diluent, 12 mL 4. Lyophilized Anti-Anatoxin-a Antibody, 3 vials 5. Antibody Diluent, 12 mL 6. Empty clear and amber HDPE bottles 7. (+)Anatoxin-a Standards: 0, 0.15, 0.40, 1.0, 2.5, 5.0 ppb; 1 mL each 8. Control at 0.75 +/- 0.185 ppb, 1 mL 9. Sample Diluent (10X) Concentrate, 2 X 25 mL 10. Wash Buffer (5X) Concentrate, 100 mL 11. Substrate (Color) Solution (TMB), 12 mL 12. Stop Solution, 12 mL