HOXA1/B1/D1 Transcription Factor Activity Assay

The HOXA1/B1/D1 Transcription Factor Activity Assay from Assay Genie quantifies the DNA-binding activity of three paralogous Hox proteins in a single 96 Assays workflow. The microplate is pre-coated with double-stranded oligonucleotides carrying the consensus Hox binding motif, so only transcription factors capable of recognizing that site in nuclear lysates are captured and reported.

Each well is incubated with your nuclear or whole-cell lysate, followed by an antibody cocktail specific for HOXA1, HOXB1, and HOXD1, then an HRP-conjugated secondary reagent. A colorimetric substrate reaction produces an absorbance signal proportional to the active fraction of these transcription factors in the input sample. The antibody panel is validated for human, mouse, and rat reactivity, allowing cross-species comparisons of Hox-driven transcriptional programs in developmental, stem cell, and cancer research models.

For best results, prepare nuclear extracts from freshly harvested cells or aliquots stored frozen at -80 degrees C, and avoid repeated freeze-thaw cycles which erode DNA-binding activity. A typical reaction uses approximately 10 uL of nuclear extract per well, with the supplied assay buffer, wash buffers, and developing reagents sufficient for the full microplate run. Researchers commonly apply this kit to study HOXA1 in hindbrain patterning, HOXB1 in motor neuron specification, and HOXD1 in limb and skeletal development.