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Nonradioactive gel mobility shift assay using chemiluminescent detection

Berger, R.; Duncan, M.R.; Berman, B.

Biotechniques 15(4): 650 652

1993

ISSN/ISBN: 0736-6205
PMID: 8251167
Document Number: 420964
A nonradioactive gel mobility shift assay using chemiluminescent detection following semidry transfer to nylon membranes is described. The procedure utilizes digoxigenin-labeled oligonucleotides in conjunction with anti-digoxigenin antibody Fab fragments coupled to alkaline phosphatase. Detection of alkaline phosphatase is by autoradiography of the chemiluminescence produced during enzymatic dephosphorylation of a dioxetane substrate. This method offers similar sensitivity to radioactive methods while having the advantages of multiple exposures, faster processing, stability of labeled oligonucleotides and safety associated with nonradioactive methods of detection.

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