TransAM® kits are sensitive, non-radioactive transcription factor ELISA-based assays that facilitate the study of transcription factor activation in mammalian tissue and cell extracts. These optimized assays are available for many different transcription factors, outlined in the table below.
How TransAM Transcription Factor ELISAs Work
The TransAM format is perfect for assaying transcription factor binding to a consensus-binding site. TransAM Kits contain a 96-stripwell plate to which the consensus-binding site oligo has been immobilized. Activated nuclear extract is added to each well and the transcription factor of interest binds specifically to this bound oligonucleotide. A primary antibody specific for an epitope on the bound and active form of the transcription factor is then added followed by subsequent incubation with secondary antibody and Developing Solution to provide an easily quantified, sensitive colorimetric readout (Figure 1).
Figure 1: Flow chart of the TransAM process.
Advantages of TransAM Transcription Factor ELISAs
Historically, transcription factor studies have been conducted using EMSAs/gel shift assays, western blots, and reporter-based assays with plasmid transfections, which are all time-consuming, do not allow for high-throughput analysis and provide only semi-quantitative results.
TransAM assays are up to 100 times more sensitive than gel shifts and can be completed in fewer than 5 hours. The TransAM assay is a non-radioactive ELISA-based assay, making them easier to perform. The high-throughput stripwell format enables simultaneous screening of up to 96 samples. Inconsistencies due to variable reporter plasmid transfection efficiencies are also eliminated, along with the need to construct stable cell lines.
Why Use TransAM Over Traditional Methods?
- Sensitivity: Up to 100-fold more sensitive than gel shift assays
- Convenience: Eliminates the use of radioactivity and the need to run gels
- It's Fast: Results can be obtained in fewer than 5 hours
- Ease of Analysis: Colorimetric readout enables simple quantitative analysis with spectrophotometry at 450 nm
- Flexible Format: 96-stripwell format enables both high- and low-throughput assays
Our TransAM Assay kits have been cited in more than 1,000 publications.
For more information about individual TransAM kits, or to find out additional information about the publication history of each kit, please click on the target of interest in the table below.
|Available Transcription Factor Assays|
|CREB & pCREB||Elk-1|
|PPARγ||Sp1 and Sp1/Sp3|
|STAT3 and STAT Family||T-bet|