c-Fos Transcription Factor Assay Kit (Colorimetric) (ab207194)
Key features and details
- Assay type: Semi-quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 3 hr 30 min
- Sample type: Nuclear Extracts
- Sensitivity: 1250 ng/well
Properties
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Storage instructions
Please refer to protocols. -
Components 1 x 96 tests 5 x 96 tests 10X Antibody Binding Buffer 1 x 2.2ml 1 x 11ml 10X Wash Buffer 1 x 22ml 1 x 110ml 96-well assay plate 1 unit 5 units Anti-rabbit HRP-conjugated IgG (0.25 μg/μL) 1 x 11µl 1 x 55µl AP-1 Mutated oligonucleotide (10 pmol/μL) 1 x 100µl 1 x 500µl AP-1 Wild-type oligonucleotide (10 pmol/μL) 1 x 100µl 1 x 5µl Binding Buffer 1 x 10ml 1 x 50ml c-Fos antibodies 1 x 11µl 1 x 55µl Developing Solution 1 x 11ml 1 x 55ml Dithiothreitol (DTT) (1 M) 1 x 100µl 1 x 500µl K-562(TPA) nuclear extract (2.5µg/μL) 1 x 40µl 1 x 200µl Lysis Buffer 1 x 10ml 1 x 50ml Plate sealer 1 unit 5 units Poly [d(l-c)] (17 µg/μL) 1 x 100µl 1 x 500µl Protease Inhibitor Cocktail 1 x 100µl 1 x 500µl Stop Solution 1 x 11ml 1 x 55ml -
Research areas
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Function
Nuclear phosphoprotein which forms a tight but non-covalently linked complex with the JUN/AP-1 transcription factor. In the heterodimer, FOS and JUN/AP-1 basic regions each seems to interact with symmetrical DNA half sites. On TGF-beta activation, forms a multimeric SMAD3/SMAD4/JUN/FOS complex at the AP1/SMAD-binding site to regulate TGF-beta-mediated signaling. Has a critical function in regulating the Has a critical function in regulating the development of cells destined to form and maintain the skeleton. It is thought to have an important role in signal transduction, cell proliferation and differentiation. -
Sequence similarities
Belongs to the bZIP family. Fos subfamily.
Contains 1 bZIP domain. -
Post-translational
modificationsPhosphorylated in the C-terminal upon stimulation by nerve growth factor (NGF) and epidermal growth factor (EGF). Phosphorylated, in vitro, by MAPK and RSK1. Phosphorylation on both Ser-362 and Ser-374 by MAPK1/2 and RSK1/2 leads to protein stabilization with phosphorylation on Ser-374 being the major site for protein stabilization on NGF stimulation. Phosphorylation on Ser-362 and Ser-374 primes further phosphorylations on Thr-325 and Thr-331 through promoting docking of MAPK to the DEF domain. Phosphorylation on Thr-232, induced by HA-RAS, activates the transcriptional activity and antagonizes sumoylation. Phosphorylation on Ser-362 by RSK2 in osteoblasts contributes to osteoblast transformation.
Constitutively sumoylated by SUMO1, SUMO2 and SUMO3. Desumoylated by SENP2. Sumoylation requires heterodimerization with JUN and is enhanced by mitogen stimulation. Sumoylation inhibits the AP-1 transcriptional activity and is, itself, inhibited by Ras-activated phosphorylation on Thr-232. -
Cellular localization
Nucleus. - Information by UniProt
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Alternative names
- Activator protein 1
- AP 1
- C FOS
see all -
Database links
- Entrez Gene: 2353 Human
- Entrez Gene: 14281 Mouse
- Entrez Gene: 314322 Rat
- Omim: 164810 Human
- SwissProt: P01100 Human
- SwissProt: P01101 Mouse
- SwissProt: P12841 Rat
- Unigene: 246513 Mouse
see all
Images
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Nuclear extracts from K-562 cells stimulated with TPA were assayed for activity of AP1 family member c-Fos.
Nuclear extracts from K-562 cells stimulated with TPA (Gray) were tested for activity of AP1 family member c-Fos with 5 µg/well of nuclear extract in the absence or presence of wild-type (Black) or mutated (White) consensus binding oligonucleotides. These results are provided for demonstration purposes only.
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Different amounts of unstimulated (gray) and PMA/Ionomycin-stimulated (black) WI-38 cells were tested for AP1 activation. These results are provided for demonstration purposes only.Different amounts of unstimulated (gray) and PMA/Ionomycin-stimulated (black) WI-38 cells were tested for AP1 activation. These results are provided for demonstration purposes only.
