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® NFAT Transcription Factor ELISA Kits
TransAM NFAT Kits provide everything needed to study Nuclear Factor of Activated T-cells, cytoplasmic, calcineurin-dependent 1 (NFATc1), including a positive control extract. The kit can be used with human and mouse extracts. See the NFAT Info tab below for kit data and more information; the kit manual can be downloaded under the Documents tab.
NFAT Transcription Factor Info
The Nuclear Factor of Activated T cells (NFAT) family of transcription factors contains five members (NFAT 1-5). The NFATC transcription complex consists of two components: a preexisitng cytosolic component that translocates to the nucleus and an inducible nuclear component. NFATc1, also known as NFATC or NFAT2, has multiple isoforms as a result of alternative splicing, that regulate inducible expression of cytokine genes in T-cells. NFATc1 is confined to the cytoplasm in resting cells, where it maintains its phosphorylation state by the action of constitutive kinases. Upon stimulation, NFATc1 is dephosphorylated by calcineurin and migrates to the nucleus. NFATc1 can be stimulated by T cell receptor (TCR) stimulation and calcium ionophores. Cyclosporin A and FK506 immunosuppressor drugs inhibit calcineurin activity on NFAT.
Figure 1: Monitoring NFATc1 activation with the TransAM NFATc1 Kit.
Different amounts of nuclear extract from untreated Raji and HeLa cells are tested for NFATc1 activation using the TransAM NFATc1 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,