IRF Transcription Factor Info
Interferon Regulatory Factors (IRFs) are a family of transcription factors involved in host defense via regulation of anti-viral immune responses, cell growth and hematopoietic development. IRF-3 is inactive in its cytoplasmic form, but upon phosphorylation by serine/threonine kinases it translocates to the nucleus and binds CREB binding protein (CREBBP) to form a complex known as dsRNA-activated factor 1 (DRAF1). This complex activates the transcription of interferon-induced genes under the control of the interferon-stimulated response element (ISRE). IRF-7 is activated by a phosphorylation event in response to infection, which leads to the activation of virus-inducible cellular genes.
Figure 1: Monitoring IRF activation with the TransAM IRF-3 (Human) Kit.
0.6 to 10 µg of poly (I/C) treated COS-7 cells were tested for IRF-3 activity using the TransAM IRF-3 (Human) Kit.
Figure 2: Monitoring IRF activation with the TransAM IRF-7 Kit.
40 ng to 0.6 µg of IRF-7 transfected COS-7 cells were tested for IRF-7 activity using the TransAM IRF-7 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 of 1-96 samples. Inconsistencies due to variable reporter plasmid transfections are eliminated, along with the need to construct stable cell lines.
Why use TransAM® transcription factor ELISAs?
- Up to 100-fold more sensitive than gelshift assays
- Eliminates the use of radioactivity and the need to run gels
- Results in less than five hours
- Colorimetric readout enables easy, quantitative analysis with spectrophotometry at 450 nm
- 96-stripwell format enables both high and low throughput
How TransAM® transcription factor ELISAs work
The TransAM format is perfect for assaying transcription factor binding to a co