Anti-Aprotinin IgG Antibodies ELISA
The Anti-Aprotinin IgG Antibodies ELISA is For Research Use Only
Size: 1×96 wells
Sensitivity: 12.5 U/ml
Dynamic Range: 12.5 U/ml – 200 U/ml
Incubation Time: 2 hours
Sample Type: Serum
Sample Size: 5 µL
Aprotinin is used in operations to reduce intra-operative and postoperative bleeding tendency. Aprotinin as an allogeneic protein has antigenic properties. It has been shown that formation of IgG antibodies to aprotinin takes place in about 50% of all treated patients. During aprotinin reexposure allergic reactions may occur due to these preformed anti-aprotinin antibodies. Lethal anaphylactic shocks have been described in literature. This Anti-Aprotinin Antibodies Assay Kit allows the detection of IgG antibodies against aprotinin in human serum.
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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.
- Prepare all reagents and samples as directed in the previous section.
- Pipette 100 µl of diluted samples, standards, controls or diluent (as blank) into the wells.
- Seal wells with adhesive strip and incubate for 1 hour at room temperature.
- 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.
- Dispense 100 µl of diluted HRP conjugate into each well
- Seal wells with adhesive strip and incubate for 30 minutes at room temperature.
- Repeat the wash as in step 4.
- Dispense 100 µl of TMB substrate solution into each well.
- Incubate for 15 minutes at room temperature in the dark.
- Add 100 µl of stop solution to each well.
- Determine the absorbance within 30 minutes at 450 nm. A reference wavelength of 620 nm/690 nm is recommended.
Typical Standard Curve