Assay Principle


HMGB1 ELISA Kit II quantifies HMGB1 by sandwich ELISA. 10 µL specimen is added to immobilized anti-HMGB1 antibody on the well together with 100 µL sample diluent, allowing HMGB1 to specifically bind to the antibody. Peroxidase (POD)-conjugated secondary antibody is then added to the sample well. Chromogenic solution is added to the antigen-antibody complex for assaying absorbacen at 450 nm.

Assay Outline

Add 100 µL Sample Diluent to each well.
Add 10 µL samples to each well.
↓ Incubate for 20-24 hours at 37°C
Wash wells 5 times.
Add 100µL POD-conjugate solution to each well.
↓ Incubate for 2 hours at 25°C
Wash wells 5 times.
Add 100µL substrate solution to each well.
↓ Incubate for 30 minutes at room temperature.
Add 100µL stop solution to each well.
Read at 450 nm.

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