Anti-Aprotinin IgG Antibodies ELISA

Additional Information

Assay Principle

Aprotinin has been pre-coated onto a microtiter plate. During incubation the anti-aprotinin antibodies are immobilized on the plate. For detection of bound antibodies an enzyme-linked anti-human-IgG anti­body conjugate is added. After washing off any unbound conjugate, a substrate solution is added. The color developing correlates to the amount of bound antibody conjugate. Absorption at 450 nm is proportional to the concentration and/or avidity of anti-aprotinin antibodies.

Assay Procedure

1. Prepare all reagents and samples as directed in the previous section.
2. Pipette 100 µl of diluted samples, standards, controls or diluent (as blank) into the wells.
3. Seal wells with adhesive strip and incubate for 1 hour at room temperature.
4. Aspirate fluid from wells and wash three times with 300 µl wash buffer. After the last wash, invert the plate and tap on a clean paper towel.
5. Dispense 100 µl of diluted HRP conjugate into each well
6. Seal wells with adhesive strip and incubate for 30 minutes at room temperature.
7. Repeat the wash as in step 4.
8. Dispense 100 µl of TMB substrate solution into each well.
9. Incubate for 15 minutes at room temperature in the dark.
10. Add 100 µl of stop solution to each well.
11. Determine the absorbance within 30 minutes at 450 nm. A reference wavelength of 620 nm/690 nm is recommended.

Typical Standard Curve