Allergens and other pollutants in house dust are collected using a variety of dust samplers that are assumed to operate similarly. This factorial design study compared sampler performance under controlled environmental conditions. House dust with known particle sizes (212-90, 90-45, and <45 μm) and allergen concentrations were sampled from new carpet squares with varying denier, pile height and pile densities. Dust mass and allergen recovery for total dust mites (Der p 1 and Der f 1), cat (Fel d 1) and cockroach allergen (Bla g 1) were assessed using the Eureka Mighty Mite (EURK), the High Volume Small Surface Sampler (HVS), or the American Industrial Hygiene Association (AIHA) method. Allergen concentrations were quantified using enzyme-linked immunosorbent assays, and dust mass results were corrected for carpet fiber shedding. Samplers were compared by exploring mass collection efficiency (CE — fiber corrected dust mass/applied dust mass) and concentration ratio (CR = allergen concentration in collected sample/allergen concentration in test dust). Test dust allergen concentrations varied by particle size fraction due to varying laboratory performance over time. The EURK and HVS samplers had CEs of 41-63% in the small and medium particle size fractions, and collected less than 20% of the available dust from the large size fraction. The AIHA CE ranged from 10% to 17% in the medium and small particle size fractions, but collected little dust in the largest particle size fraction. The AIHA and HVS samplers were more likely to acquire more representative and less variable allergen CRs compared with the EURK method. Health studies that use allergen concentration as an exposure metric need to consider the implications of sampler performance when interpreting links to health outcomes and development of health-based standards for allergens in house dust.