The COVID-19 pandemic has provided a new work system. It is called work from home. This phenomenon continues after the COVID-19 pandemic. It gives rise to a business sector where workers can work from home. This phenomenon raises problems for men and women related to work-life balance. Therefore, the existence of new phenomena and their problems will require the development of analyses in measurement to suit the existing phenomena and problems. One is a study of item bias on the work-life balance scale based on gender and age in the context of working from home. This research aims to determine whether there was an item bias on work-life balance based on age and gender. The sample of this research amounted to 130 workers consisting of 65 female workers and 65 male workers with an age range from 19-51 years. The sampling technique used was the quota sampling technique. The analysis carried out was the Differential Item Functioning (DIF) analysis on the work-life balance scale measuring instrument. The research shows that the item has a probability value below 0.05, i.e., 0.044 in item no.4, based on gender. Furthermore, two items have a probability value below 0.05, i.e., 0.031 (item no.2) and 0.003 (item no.15), based on age. Therefore, it can be concluded that there is item bias based on gender and age.