Many causes for falls from ladders in construction are related to the user’s activities; however, the extent to which users comply with ladder use best practices is unknown and has not been well quantified before. We developed and tested an audit tool that assesses compliance with best practices guidelines for portable ladder use designed for applications in the construction industry. Implemented on a hand-held computer, the assessment tool consisted of a series of checklists categorized in four groups; ladder condition, setup, moving on a ladder, and completing tasks from a ladder. For these four observational categories, the resulting tool contained 31 and 33 questions for step and extension ladders, respectively. Three individuals trained to use the tool scored a set of photographs and videos depicting 25 ladder conditions, 20 ladder setups, 10 users moving on ladders, and 13 users completing tasks from a ladder for a total of 78 observations. The assessment tool had good agreement across and within raters. For the three raters, agreement ranged from 79% to 97% across the questions. Within one subject, kappa coefficients for the intrarater reliability ranged from 0.67 to 0.91. The tool offers a practical method to quantify best practices associated with ladder use that can ultimately inform targeted intervention efforts.