|
|
|||||||||||||||||||||||||||||
|
Transcription Factor Microplate Assays Marligen’s Chemiluminescent Transcription Factor Microplate Assays allow researchers to quantitate the DNA binding activity of transcription factors in nuclear extracts prepared from cultured cells or tissues. The assays are designed to be used in combination with Marligen’s Nuclear Extraction Kit to ensure optimal performance. Transcription factors are DNA-binding proteins that regulate gene expression through the activity of two distinct domains. One domain displays high affinity for a specific DNA sequence and the other interacts with coactivators, corepressors and/or the basal transcriptional machinery that transcribes genes. Transcription factors are activated by a variety of mechanisms including ligand binding, post-translational modifcations, and interactions with other proteins involved in the regulation of transcription. Marligen’s Chemiluminescent Transcription Factor Microplate Assays are intended for use in studying the mechanisms of gene regulation and in analyzing signaling pathways important for development, differentiation, control of cell division and death, etc. Marligen’s Chemiluminescent Transcription Factor Microplate Assays can be used in drug discovery applications including disease characterization, target validation, compound screening, and toxicity testing. Marligen’s rapidly expanding line of products for studying gene regulation also includes Multiplex Transcription Factor Assay Kits and Testing Services, Nuclear Extraction Kits and Transcription Factor Activators, Inhibitors, Proteins and Antibodies. Technology Standard methods for studying transcription factors and gene regulation include gel-shifts, western blot and reporter gene assays, which are time-consuming, do not allow for high-throughput and provide only semi-quantitative results. Marligen’s Chemiluminescent Transcription Factor Microplate Assays employ a proprietary assay technology and chemiluminescent detection that offers superior sensitivity, broad dynamic range and excellent reproducibility compared with traditional methods or with commercially available transcription factor ELISAs. The assays do not require the use of radioactivity, and results can be obtained in under four hours. The convenient stripwell format enables testing of from one to 96 samples in each plate, and the chemiluminescent signal is measured with standard 96-well plate luminometers. A schematic of the assay technology and workflow is shown in Figure 1.
Figure 1: Summary of protocol for Marligen’s Chemiluminescent Transcription Factor NFκB Microplate Assay. Features and Benefits
Figure 2: NFκB activation was observed in less than 0.1 μg of nuclear extract from TNF-α stimulated HeLa cells
Figure 3: Marligen’s Chemiluminescent AP-1 Microplate Assay detects both c-jun homodimers and c-jun-c-fos heterdimers, obviating the need for specific antibodies to multiple protein forms or splice variants.
Figure 4: Changes in the activity of multiple transcription factors are observed when 10 μg of nuclear extract from PMA treated cells are tested on Marligen’s 20-plex assay (Figure 4A). Single Analyte Chemiluminescent assays require less sample and can be used to further characterize individual responses, Figure 4B shows data from PPAR assay. Please click here for a description of other products and services offered by Marligen. |
