Transcription Factor Assays
DNA-binding ELISA kits that semi-quantitatively detect active transcription factors — NF-κB, p53, CREB, SMAD, STAT, c-Jun, Myc and 100+ more — directly from nuclear or cell lysates, with no radioactivity and no gel shift.
Measure DNA-binding across 96 wells at once.
Far faster than EMSA, gel shift or western blot workflows.
No radioactive labelling required.
Run many samples and treatments in parallel.
Total-activity and phospho-specific assays for 100+ factors.
An indirect ELISA: biotinylated consensus DNA is captured on a streptavidin-coated plate, the active transcription factor in your lysate binds its DNA motif, and a specific primary antibody plus HRP secondary produce a colour signal proportional to the active factor.
Wells are pre-coated with streptavidin.
Biotinylated consensus oligonucleotides bind via biotin–streptavidin.
Active transcription factor binds its DNA motif.
Primary antibody and HRP secondary bind the captured factor.
TMB turns blue, then yellow on stopping; signal ∝ active TF.
Semi-quantify transcription factor DNA-binding in nuclear and cell lysates.
Analyse activation of transcription factors across signalling pathways.
Investigate candidate targets in drug-discovery programmes.
Screen compounds for modulators of transcription factor activity.
Establish how phosphorylation drives transcription factor activation.
All 201 total-activity and phospho-specific DNA-binding assays, grouped by transcription factor family.
It measures the DNA-binding activity of a specific transcription factor. Only the active factor binds the immobilised consensus DNA sequence, so the signal reflects activation rather than total protein level. Results are semi-quantitative, for comparing samples and treatments.
Nuclear extracts or whole-cell lysates. Nuclear extracts usually give the strongest signal because active transcription factors accumulate in the nucleus.
Total-activity assays detect all DNA-bound forms of the factor. Phospho-specific assays use an antibody to a defined phosphorylation site, so you can see how a specific phosphorylation event relates to DNA binding.
It is most likely still usable, though activity may be slightly reduced. The most temperature-sensitive components are the standards, HRP conjugate and detection antibody.
Common causes are an unsuitable nuclear lysate, poor lysate preparation or storage, or missed steps. Add protease and phosphatase inhibitors, keep everything on ice, store lysates at −80°C avoiding freeze–thaw, and follow the manual closely for reagent volumes.
Usually inadequate or uneven washing, too much antibody, lysate or substrate, over-long substrate incubation, or contamination. Wash thoroughly with the correct volumes, reduce antibody, lysate or substrate as needed, and check your multichannel pipette or plate washer is calibrated.
Our PhD-level team can develop custom DNA-binding assays on request — ask about a custom assay.