You will need
A washing bottle, a graduated cylinder, a ScreenShaker, a comb (trough holder), an incubation box, a 1OO µl - 1000 µl pipette, a timer and some gloves.
Sera and all solutions must be at ambient temperature (20°C - 22°C).
Dilute the washing buffer concentrate (1:25).
Processing the test
(1) Wet the membranes with washing buffer. Remove the washing buffer by gently tapping the troughs on a paper towel. The membranes should be wet but with no surplus buffer on its surface.
(2) Pipette 250 µl serum (AllergyScreen®) or 300 µl serum (AlleisaScreen®) into the troughs, taking care that the membranes are completely covered. Carefully silde the comb into the incubation box. Place the incubation box onto the ScreenShaker. Incubate for 45 minutes at ambient temperature.
(3) Carefully take the comb out of the box. Remove the sera by rinsing the troughs with the washing buffer. Rinse each membrane five times, thereby shake the washing buffer in the troughs for some seconds. Remove the surplus by gently tapping the comb on paper towels.
(4) Pipette 250 µl (AllergyScreen®) or 300 µl (AlleisaScreen®) detection antibody in each trough. Slide the comb into the incubation box and incubate for another 45 minutes on the ScreenShaker at ambient temperature.
(5) Wash the membranes as described in step (3).
(6) Pipette 250 µl (AllergyScreen®) or 300 µl (AlleisoScreen®) of the streptavidin conjugate into each reaction trough. Replace the comb into the incubation box and incubate for exactly 20 minutes on the ScreenShaker.
(7) Wash the membraries as described in step (3), but this time rinse each membrane 10 times.
(8) Pipette 250 µl (AllergyScreen®) or 300 µl (AlleisaScreen®) of the substrate into each reaction trough. Slide the comb into the incubation box and replace it on the ScreenShaker. Incubate for exactly 20 minutes at ambient temperature on the incubation shaker.
(9) Rinse each membrane several times under running water. Remove the washing buffer by gently tapping the through on a paper towel. Leave the membranes to air dry or use a conventional hair dryer.
The background of the membranes will turn white, while the control line and positive tested allergens will stay purple.