GATA (GATA1, GATA2, GATA3) Transcription Factor Assay Kit (Colorimetric) (ab207205)
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 GATA 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 GATA-1 antibody 1 x 11µl GATA-2 antibody 1 x 11µl GATA-3 antibody 1 x 11µl Herring sperm DNA (1 μg/μL) 1 x 100µl K-562 nuclear extract (5 μg/μL) 1 x 40µl Lysis Buffer 1 x 10ml Mutated oligonucleotide (10 pmol/μL) 1 x 100µl Plate sealer 2 units Protease Inhibitor Cocktail 1 x 100µl Stop Solution 1 x 60ml Wild-type oligonucleotide (10 pmol/μL) 1 x 100µl -
Research areas
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Function
Transcriptional activator or repressor which probably serves as a general switch factor for erythroid development. It binds to DNA sites with the consensus sequence 5'-[AT]GATA[AG]-3' within regulatory regions of globin genes and of other genes expressed in erythroid cells. Activates the transcription of genes involved in erythroid differentiation of K562 erythroleukemia cells, including HBB, HBG1/2, ALAS2 and HMBS (PubMed:24245781). -
Tissue specificity
Erythrocytes. -
Involvement in disease
X-linked dyserythropoietic anemia and thrombocytopenia
Thrombocytopenia with beta-thalassemia, X-linked
Anemia without thrombocytopenia, X-linked -
Sequence similarities
Contains 2 GATA-type zinc fingers. -
Domain
The two fingers are functionally distinct and cooperate to achieve specific, stable DNA binding. The first finger is necessary only for full specificity and stability of binding, whereas the second one is required for binding. -
Post-translational
modificationsHighly phosphorylated on serine residues. Phosphorylation on Ser-310 is enhanced on erythroid differentiation. Phosphorylation on Ser-142 promotes sumoylation on Lys-137.
Sumoylation on Lys-137 is enhanced by phosphorylation on Ser-142 and by interaction with PIAS4. Sumoylation with SUMO1 has no effect on transcriptional activity.
Acetylated at 2 conserved lysine-rich motifs by CREBBP in vitro. Acetylation does not affect DNA-binding in vitro but is essential to induce erythroid differentiation and for binding chromatin in vivo (By similarity). Acetylated on Lys-233, Lys-245 Lys-246 by EP300. -
Cellular localization
Nucleus. - Information by UniProt
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Alternative names
- Eryf1
- Erythroid transcription factor
- GATA 2
see all -
Database links
- Entrez Gene: 2623 Human
- SwissProt: P15976 Human
- SwissProt: P23769 Human
- SwissProt: P23771 Human
- Unigene: 765 Human
Images
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Typical Data
Nuclear extracts (5 µg/well) were tested for GATA activation: K-562 cells were used for GATA-1 and GATA-2 detection and Jurkat cells were used for GATA-3 detection. Activation was monitored in absence (grey) and in the presence of wild-type (black) or mutated (white) consensus binding oligonucleotides. Note that the wild-type oligonucleotide reduces GATA binding by over 90%, while incubation with the mutant GATA competitor oligo has a limited effect on GATA-1, -2 and -3 binding to DNA. These results are provided for demonstration purposes only.