Recombinant Human Homeobox protein Hox-D10 (HOXD10) is a sequence-specific transcription factor that belongs to the homeobox - containing transcription factor family. HOXD10 protein plays crucial roles in various biological processes, especially during embryonic development.It is involved in patterning and differentiation of tissues and organs, particularly in the development of the axial skeleton and limb buds.It has also been reported to suppress ovarian cancer, hepatocellular carcinoma and other cancer cells. Re-expression of HOXD10 results in significant inhibition of cell survival, induction of cell apoptosis and impairment of cell migration and invasion. Besides,Hoxd-10?can?bind?to?and?transactivate?the?Shh?promoter?in?vitro, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human HOXD10 and recombinant human SHH . Briefly, biotin-linked HOXD10 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to SHH-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50μl stop solution to the wells and read at 450nm immediately. The binding activity of HOXD10 and SHH was shown in Figure 1, the EC50?for this effect is 0.023ug/mL.