The influence of β-glucans on the properties of gluten-free dough and bread is still not fully explained, with the literature suggesting both positive and negative effects. The aim of this study was to investigate the effect of the molar mass of oat β-glucans on the properties of gluten-free bread. Gluten-free breads were baked under standardized conditions from a model gluten-free mix without and with a 1% or 2% share of oat β-glucans of a low molar mass of 24,540 g/mol, a medium molar mass of 85,940 g/mol and a high molar mass of 1,714,770 g/mol. The share of β-glucans affected the increase in water addition to the baking mix and dough yield proportionally to the molar mass and amount of β-glucans. The β-glucans of the highest molar mass, particularly at a 2% share, were most effective in increasing bread volume, reducing hardness and increasing the moisture content of the bread crumb on the day of baking, as well as reducing the increase in hardness and maintaining a high moisture content of the bread crumb after 1 day of storage, compared to bread without added β-glucans.