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® C/EBP ?/? Transcription Factor ELISA Kits
TransAM C/EBP (CCAAT-enhancer binding protein) Kits provide everything needed to study activated C/EBP? or C/EBP?, including a positive control extract. The C/EBP ?/? kit can be used with human, mouse and rat extracts. Click the C/EBP Info tab below for data and more information; kit manuals can be downloaded under the Documents tab.
C/EBP Transcription Factor Info
C/EBPs plays an important role in regulating the balance between cell growth and differentiation. The family of C/EBP (CCAAT-enhancer binding protein) basic leucine zipper transcription factors includes C/EBP?, C/EBP?, C/EBP?, C/EBP? and C/EBP?. Each member exhibits similar DNA-binding specificities and contains a leucine zipper dimerization region. C/EBPs can bind DNA as homodimers and recruit coactivators to promote gene expression. C/EBP? can also heterodimerize with C/EBP? and C/EBP?. The TransAM C/EBP ?/? Kit contains antibodies specific for the active form of C/EBP? and C/EBP? when bound to the target DNA.
Figure 1: Specificity of C/EBP? binding.
Increasing amounts of rat liver nuclear extract are assayed using the TransAM C/EBP ?/? Kit in the presence or absence of competitor oligonucleotides containing the wild-type or a mutated form of the C/EBP consensus binding site.
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 fo