STAT (STAT1alpha, STAT3, STAT5A, STAT5B) Transcription Factor Assay Kit (Colorimetric) (ab207228)
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: 600 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 IRF-3 assay plate 2 units Anti-rabbit HRP-conjugated IgG (0.25 μg/μL) 2 x 11µl Binding Buffer 1 x 10ml Developing Solution 2 x 11ml Dithiothreitol (DTT) (1 M) 1 x 100µl Herring sperm DNA (1 μg/μL) 1 x 100µl Lysis Buffer 1 x 10ml Nb nuclear extract (prolactin stimulated) (2.5 μg/μL) 1 x 40µl Plate sealer 2 units Protease Inhibitor Cocktail 1 x 100µl STAT mutated oligonucleotide (10 pmol/μL) 1 x 100µl STAT Wild-type oligonucleotide (10 pmol/µL) 1 x 100µl STAT1α antibody 1 x 11µl STAT3 antibody 1 x 20µl STAT5A antibody 1 x 11µl STAT5B antibody 1 x 11µl Stop Solution 1 x 60ml -
Research areas
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Cellular localization
STAT3: Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm. Constitutive nuclear presence is independent of tyrosine phosphorylation. Predominantly present in the cytoplasm without stimuli. Upon leukemia inhibitory factor (LIF) stimulation, accumulates in the nucleus. The complex composed of BART and ARL2 plays an important role in the nuclear translocation and retention of STAT3. STAT 5A + B: Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation. -
Database links
- Entrez Gene: 600555 Human
- Entrez Gene: 6774 Human
- Entrez Gene: 6776 Human
- Entrez Gene: 6777 Human
- Entrez Gene: 20846 Mouse
- Entrez Gene: 20848 Mouse
- Entrez Gene: 20850 Mouse
- Entrez Gene: 20851 Mouse
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Images
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Different amounts of Nb2 were tested for STAT Family activation.
Nuclear extracts from Nb2 cells stimulated with prolactin) were assayed for activity of STAT family members: STAT1alpha, STAT3, STAT5A, STAT5B in the absence (grey) or presence of wild-type (black) or mutated (white) consensus binding oligonucleotides. Note that the wild-type oligonucleotide reduces STAT binding by over 90% while incubation with the mutant STAT competitor oligo has limited effect on STAT binding to DNA. These results are provided for demonstration purposes only.