NONRADIOACTIVE GEL MOBILITY SHIFT ASSAY USING CHEMILUMINESCENT DETECTION

A nonradioactive gel mobility shift assay using chemiluminescent detec tion following semidry transfer to nylon membranes is described. The p rocedure utilizes digoxigenin-labeled oligonucleotides in conjunction with anti-digoxigenin antibody Fab fragments coupled to alkaline phosp hatase. Detection of alkaline phosphatase is by autoradiography of the chemiluminescence produced during enzymatic dephosphorylation of a di oxetane substrate. This method offers similar sensitivity to radioacti ve methods while having the advantages of multiple exposures, faster p rocessing, stability of labeled oligonucleotides and safety associated with nonradioactive methods of detection.