TransAM® 転写因子活性測定ELISA

sensitive, quantitative ELISAs for transcription factor activation

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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.

View Available Transcription Factors

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).

Flow chart of the TransAM DNA binding transcription factor ELISA method for measurement of activated transcription factors
Figure 1: Flow chart of the TransAM process.

Activated transcription factor in the cell extract binds to the consensus-binding site on the oligo immobilized in the well. Incubation with the supplied primary and secondary antibodies specifically quantifies the amount of activated transcription factor.

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
AP-1 ATF-2
c-Myc C/EBP α/β
CREB & pCREB Elk-1
ER FKHR (FOXO1)
GATA GR
HIF-1 HNF
IRF-3 MAPK Family
MEF2 MyoD
NF-YA NFATc1
NFκB Nrf2
Oct-4 p53
PPARγ Sp1 and Sp1/Sp3
STAT3 and STAT Family T-bet

 

Technology covered under EAT-filed patents and licensed to Active Motif. Use of TransAM in NFκB-related drug discovery may be covered under U.S. Patent No. 6,150,090 and require a license from Ariad Pharmaceuticals (Cambridge, MA, USA).