Test procedure
Binding of Staph. Enterotoxins present in the sample
• add 100 μl of controls or samples, respectively, to each well
• incubate at 35 - 37 °C for 1 h (cover plate)
First washing
• wash each well 5 times with 300 μl washing buffer
Adding Conjugate 1
• add 100 μl conjugate 1 to all wells
• incubate at 35 - 37 °C (95 - 99 °F) for 1 h (cover plate)
Second washing
• wash each well 5 times with 300 μl washing buffer
Adding Conjugate 2
• add 100 μl conjugate 2 to all wells
• incubate at 35 - 37 °C (95 - 99 °F) for 30 min
Third washing
• wash each well 5 times with 300 μl washing buffer
Adding Substrate/Chromogen and reading
• add 100 μl substrate/chromogen to each well
• incubate 15 min at 35 - 37 °C (95 - 99 °F) in the dark
• colour change from red to blue indicates positive samples
Adding Stop Solution and reading
• add 100 μl stop solution
• read absorbance at 450 nm with a microtiter photometer