In dairy herds, application of antimicrobials at drying-off is a common mastitis control measure. This article describes a protocol for systematic review and meta-analysis to address three crucial points regarding antimicrobial usage at drying-off: (1) comparative efficacy of antimicrobials used for preventing new and eliminating existing intramammary infections (IMI); (2) comparison of selective and blanket dry cow therapy approaches in preventing new and eliminating existing IMI; and (3) assessment of the extra prevention against new IMI that can be gained from using antimicrobial-teat sealant combinations versus antimicrobials alone. Five PICO (Population, Intervention, Comparator, Outcome) questions were formulated to cover the three objectives of the review. Medline, CAB Abstracts, Web of Science, and conference proceedings will be searched along with iterative screening of references. Articles will be eligible if: (1) published after 1966; (2) written in English or French; and (3) reporting field clinical trials and observational studies, conducted on dairy cows at drying-off, with at least one antimicrobial-treated group and one IMI-related outcome. Authors will independently assess the relevance of titles and abstracts, extract data, and assess bias and the overall quality of evidence. Results will be synthesized and analyzed using pairwise and network meta-analysis. The proposed study will significantly update previously conducted reviews.