TransAM® C/EBP α/β
dna-binding ELISA for activated C/EBP α and C/EBP β transcription factors
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 α/β Transcription Factor ELISA Kits
TransAM C/EBP (CCAAT-enhancer binding protein) Kits provide everything needed to study activated C/EBPα or C/EBPβ, including a positive control extract. The C/EBP α/β 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.
|TransAM® C/EBP α/β||1 x 96 rxns||44196||$665||Buy Now|
|5 x 96 rxns||44696||$2,795||Buy Now|
|TransAM® C/EBP α/β Manual|
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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/EBPα, C/EBPβ, C/EBPδ, C/EBPγ and C/EBPε. 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/EBPα can also heterodimerize with C/EBPβ and C/EBPγ. The TransAM C/EBP α/β Kit contains antibodies specific for the active form of C/EBPα and C/EBPβ when bound to the target DNA.
Figure 1: Specificity of C/EBPβ binding.
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 consensus-binding site. TransAM Kits contain a 96-stripwell plate to which the consensus-binding site oligo has been immobilized. Activated nuclear extract is added to each well and the transcription factor of interest binds specifically to this bound oligonucleotide. A primary antibody specific for an epitope on the bound and active form of the transcription factor is then added followed by subsequent incubation with secondary antibody and Developing Solution to provide an easily quantified, sensitive colorimetric readout (Figure 1).
Figure 1: Flow chart of the TransAM process.
Contents & Storage
One or five C/EBP α/β 96-well plate(s) with plate sealer(s), C/EBPα and C/EBPβ primary antibodies, HRP-conjugated secondary antibody, C/EBP wild-type and mutated oligonucleotides, positive control cell extract, DTT, Herring Sperm DNA, Protease Inhibitor Cocktail, Lysis, Binding, 10X Washing and 10X Antibody Binding Buffers, and Developing and Stop Solutions. Reagent storage conditions vary from room temperature to -80°C, see manual for details. All reagents are guaranteed stable for 6 months when stored properly.