In addition to its role as an essential pollinator, the Western honeybee, Apis mellifera, is regarded as a good model insect due to its highly evolved sociality, labor division, and flexibility of colony management. Therefore, to identify the molecular mechanisms involved in various colony compositions and flexibility of worker division, it is essential to investigate the gene expression in various developmental stages of the honeybee. For target gene quantification, quantitative realtime PCR (qRT-PCR) has been widely used. However, a reliable reference gene stably expressing across different samples should be selected. Thus, to identify credible reference genes in honeybee colonies, we evaluated five candidate genes (RPS5, RPS18, GAPDH, ARF1, and RAD1a) in samples collected from seven developmental stages (egg, 1st instar larvae, 3rd instar larvae, 5th instar larvae, pupa, nurse, and forager) using three analysis programs algorithms (geNorm, NormFinder, and BestKeeper). Although the reference gene ranked slightly different in analysis algorithms, our study suggests that RAD1a is the most suitable reference gene for accurate normalization of target gene expression at the developmental stage of the honeybee.