STAT1 Transcription Factor Activity Assay

The STAT1 Transcription Factor Activity Assay from Assay Genie uses a microplate whose wells are pre-coated with double-stranded oligonucleotides containing the STAT1-specific binding motif (GAS or ISRE consensus element). Activated STAT1 — typically phosphorylated on tyrosine 701 — present in nuclear or whole-cell lysates binds these immobilized DNA sequences during incubation. A STAT1-specific primary antibody and an HRP-conjugated secondary reagent are added sequentially, and the chromogenic substrate produces a measurable signal on a standard plate reader.

This DNA-binding immunoassay is for research use only and is commonly applied to interrogate the JAK-STAT signaling axis, interferon (IFN-α and IFN-γ) responses, cytokine-driven transcription, viral infection models, and tumor immunology. Reactivity has been validated against human, mouse, and rat samples, enabling cross-species pathway studies. Nuclear extracts are typically prepared from cultured cells or primary tissue, and the kit is compatible with both fresh and frozen lysate preparations.

The workflow generally takes 4-5 hours from sample loading to final readout. Reagents are stable when stored refrigerated at 4°C, and the antibody components should be aliquoted and frozen at -20°C to minimize freeze-thaw cycles and preserve shelf life. Each kit provides reagents in a microplate format along with positive and negative controls to confirm run-to-run reproducibility. The colorimetric readout allows quantification of STAT1 DNA-binding activity relative to the supplied positive control.