AP1 (c-Fos/FosB/Fra1/c-Jun/JunD) Transcription Factor Assay Kit (Colorimetric) (ab207196)
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 2 x 96 tests 10X Antibody Binding Buffer 2 x 2.2ml 10X Wash Buffer 1 x 60ml 96-well assay plate 2 units Anti-rabbit HRP-conjugated IgG (0.25 μg/μL) 2 x 11µl AP-1 Mutated oligonucleotide (10 pmol/μL) 1 x 100µl AP-1 Wild-type oligonucleotide (10 pmol/μL) 1 x 100µl Binding Buffer 1 x 10ml c-Fos antibodies 1 x 11µl Developing Solution 2 x 11ml Dithiothreitol (DTT) (1 M) 1 x 100µl FosB antibodies 1 x 11µl Fra-1 antibodies 1 x 11µl JunB antibodies 1 x 11µl JunD antibodies 1 x 11µl K-562(TPA) nuclear extract (2.5µg/μL) 1 x 40µl Lysis Buffer 1 x 10ml Phospho-c-Jun antibody 1 x 22µl Plate sealer 2 units Poly [d(l-c)] (17 µg/μL) 1 x 100µl Protease Inhibitor Cocktail 1 x 100µl Stop Solution 1 x 60ml -
Research areas
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Cellular localization
JunB: Nucleus. FRA1: Nucleus. c-Fos+FosB+c-Jun+JunD: Nucleus. Endoplasmic reticulum. Cytoplasm, cytosol. In quiescent cells, present in very small amounts in the cytosol. Following induction of cell growth, first localizes to the endoplasmic reticulum and only later to the nucleus. Localization at the endoplasmic reticulum requires dephosphorylation at Tyr-10 and Tyr-30. -
Database links
- Entrez Gene: 2353 Human
- Entrez Gene: 3726 Human
- Entrez Gene: 8061 Human
- Omim: 136515 Human
- Omim: 165161 Human
- SwissProt: P01100 Human
- SwissProt: P05412 Human
- SwissProt: P15407 Human
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Images
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Nuclear extracts from K-562 cells stimulated with TPA were assayed for activity of AP1 family members c-Jun, c-Fos, FosB, JunB, JunD and Fra-1.
Nuclear extracts from K-562 cells stimulated with TPA (5 µg) were assayed for activity of AP1 family members c-Jun, c-Fos, FosB, JunB, JunD and Fra-1 in the absence (grey) or presence of wild-type (black) or mutated (white) consensus binding oligonucleotides. These results are provided for demonstration purposes only
