Protocol ID: Protocol Tertiary step
Title
:
Protocol Tertiary step
Category
:
Lectin-Glycan Binding
Overview
:
Unlabeled sample is diluted to an appropriate concentration in standard binding buffer (TSM, 1% BSA and 0.05% Tween 20). Seventy microliters is applied to the printed surface of the array, cover slip is applied, and incubated at room temperature in a humidified chamber away from light for one hour. The cover slip was removed and the slide is rinsed 4 times in standard TSM washing buffer containing 0.05 percent Tween and 4 times in TSM buffer without Tween. A second step is performed with seventy microliters of antibody/reagent specific for the unlabeled sample diluted in TSM binding buffer is added to the slide for one hour at room temperature in a humidified chamber away from light as described. Washes were performed as above. A tertiary step is performed with seventy microliters of fluorescently labeled antibody/reagent to the antibody/reagent in the second step, diluted in TSM buffer for one hour in a humidified chamber away from light as described. Washes are performed as above, followed by 4 washes in distilled water.
Procedure
:
See attached PDF file