TransAM® Kits are DNA-binding ELISAs that facilitate the study of transcription factor activation in mammalian tissue and cell extracts. Assays are available for over 40 different targets (see the list at right). Each kit includes a 96-stripwell plate in which multiple copies of a specific double-stranded oligonucleotide have been immobilized. When nuclear or whole-cell extract is added, activated transcription factor of interest binds the oligonucleotide at its consensus binding site and is quantified using the included antibody, which is specific for the bound, active form of the transcription factor being studied. For complete details, click the TransAM® Method tab below.
TransAM® PPAR? Transcription Factor ELISA Kits
TransAM PPAR? Kits provide everything needed to study Peroxisome Proliferator-Activated Receptor gamma (PPARg), including a positive control extract. The kit can be used to detect gamma 1 and gamma 2 from human, mouse and rat extracts. It will not cross-react with alpha or delta isotypes. See the PPAR Info tab below for kit data and more information; the kit manual can be downloaded under the Documents tab.
Peroxisome Proliferator-Activated Receptors (PPARs) are nuclear receptors involved in lipid transport and metabolism. As such, their roles in chronic diseases such as diabetes, obesity, atherosclerosis and cancer are heavily investigated. Transcriptional activity of PPARs is regulated by fatty acid binding. Three PPAR isotypes have been identified: ?, ? and ?. PPAR? stimulates lipolysis of circulating triglycerides and the subsequent uptake of fatty acids into adipose cells. PPARs bind to peroxisome proliferator response elements (PPREs) as heterodimers with the retinoid X receptor (RXR).
Figure 1: Monitoring PPAR activation with the TransAM PPAR? Kit.
Different amounts of nuclear extracts from unstimulated (opened squares) and PMA-stimulated (filled circles) THP-1 cells are tested for PPAR activation using the TransAM PPAR? Kit.
The TransAM® transcription factor ELISA advantage
Historically, transcription factor studies have been conducted using gelshift, Western blot and reporter plasmid transfections, which are time-consuming, do not allow for high-throughput and provide only semi-quantitative results. TransAM assays are up to 100 times more sensitive than gelshift techniques, and can be completed in less than 5 hours. Because TransAM is an ELISA-based assay*, there is no radioactivity, and the high-throughput stripwell format enables simultaneous screening