Human head and body lice attach their eggs respectively to human hair or clothing by female lice secreted glue that hardens into a nit sheath that protects the egg. In this study, a series of experiments were conducted to characterize the glue-like material of the nit sheath. Fourier transform infrared spectroscopy on embryo-cleared nit showed proteinaceous amide I bands. With this result, we determined the amino acid composition of the nit sheath proteins and performed similarity search against the protein products of the body louse genome to identify the candidate nit sheath proteins. The identified two homologous proteins newly named as louse nit sheath protein (LNSP) 1 and LNSP2 are composed of three domains of characteristic repeating sequences. The N-terminal and middle domains consist of tandem two-residue repeats of Gln-Ala and Gly-Ala, respectively, which are expected to fold into β-strands and may further stack into β-sheets, whereas the C-terminal domain contains multiple consecutive Gln residues. Temporal and spatial transcription profiling demonstrated that both LNSP1 and LNSP2 are most predominantly expressed in the accessory gland of females of egg-laying stage, supporting that they indeed encode the nit sheath proteins. Further adhesive property of recombinant partial LNSP1 suggests that both LNSP1 and LNSP2 may act as glues.