Product nameGATA-4 Transcription Factor Assay Kit (Colorimetric)
Sample typeNuclear Extracts
Sensitivity< 1000 ng/well
Assay time3h 30m
Species reactivityReacts with: Human
GATA-4 Transcription Factor Assay Kit (Colorimetric) (ab207206) is a high throughput assay to quantify GATA-4 activation in nuclear extracts. This assay combines a quick ELISA format with a sensitive and specific non-radioactive assay for transcription factor activation.
A specific double stranded DNA sequence containing the GATA-4 consensus binding site (5’ –AGATAA– 3’) has been immobilized onto a 96-well plate. Active GATA-4 present in the nuclear extract specifically binds to the oligonucleotide. GATA-4 is detected by a primary antibody that recognizes an epitope of GATA-4 accessible only when the protein is activated and bound to its target DNA. An HRP-conjugated secondary antibody provides sensitive colorimetric readout that at OD 450 nm. This product detects only human GATA-4.
Key performance and benefits:
- Assay time: 3.5 hours (cell extracts preparation not included).
- Detection limit: < 1 µg nuclear extract/well.
- Detection range: 1 – 10 µg nuclear extract/well.
The GATA family of transcription factors consists of 6 members: GATA 1-6. The family is defined by the presence of 1 or 2 zinc fingers containing four cysteine residues with the motif CX2CX12CX2C that coordinate a single zinc ion. In mammals, the N-finger is important for interactions with cofactors such as Friend of GATA (FOG), while the C-finger binds to (A/T)GATA(A/G) motifs. Genes encoding GATA factors are transcriptionally regulated in a tissue-restricted manner. They include platelet factor 4 (PF4), glycoprotein (GP) IIB, α-myosin heavy chain, thyroid transcription factor-1 (TTF-1) and H+/K+-ATPase. The members of the GATA family have been grouped into two subfamilies: GATA-1, -2 and -3 are expressed mainly in hematopoietic cells, and GATA-4, -5 and -6 are found predominantly in the heart and gut.
GATA-1, the founding member of the family, is essential for normal erythropoiesis and megakaryocyte differentiation. It is expressed in erythroid cells, megakaryocytes, mast cells, eosinophils and testis. Thought to be a negative regulator of cell proliferation in early megakaryocyte progenitors, GATA-1 has also been implicated in regulation of terminal differentiation markers in various myeloid cells. Mutations in GATA-1 are associated with familial blood disorders. GATA-2 is expressed in vascular endothelial cells and embryonic brain and liver. It is thought to play a role in the regulation of endothelial gene expression, and is also found in megakaryocytes and mast cells with GATA-1. GATA-3 is found mainly in cells of T-cell lineage, and in low levels in mast cells. It is detected in the most immature subset of fetal day 12 thymocytes, and is essential for T-cell development.
GATA-4, -5 and -6 regulate gene expression in the heart, gut epithelium, liver, lung and gonad.1 Target genes of GATA-4, -5 and -6 include A- and B-type natriuretic peptides, cardiac actin and α-myosin heavy chain. GATA-4 is the transcriptional activator of several cardiac muscle-specific genes and a key regulator of the cardiomyocyte gene program. It can be found in the adult heart, ovary, testis, lung, liver and small intestine. During development, GATA-4 is expressed in the heart, gut, testis, ovary, liver, visceral endoderm and parietal endoderm. GATA-5 is expressed in the allantois, heart, lung bud, urogenital ridge, bladder and gut epithelium during development, and can be found in the adult small intestine, stomach, bladder and lungs. GATA-6 is thought to regulate the proliferation of cardiac progenitor cells. During development, it is expressed in the primitive streak, allantois, visceral endoderm, heart, lung buds, urigenital ridge, vascular smooth muscle cells and epithelial layer of the stomach, small intestine and large intestine. In adults, GATA-6 is found in the heart, aorta, stomach, small intestine and bladder.
Storage instructionsPlease 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 GATA assay plate 1 unit 5 units Anti-rabbit HRP-conjugated IgG (0.25 μg/μL) 1 x 11µl 1 x 55µl Binding Buffer 1 x 10ml 1 x 50ml Developing Solution 1 x 11ml 1 x 55ml Dithiothreitol (DTT) (1 M) 1 x 100µl 1 x 500µl F9 nuclear extract (2.5 μg/μL) 1 x 40µl 1 x 200µl GATA-4 antibody 1 x 11µl 1 x 55µl Herring sperm DNA (1 μg/μL) 1 x 100µl 1 x 500µl Lysis Buffer 1 x 10ml 1 x 50ml Mutated oligonucleotide (10 pmol/μL) 1 x 100µl 1 x 500µl Plate sealer 1 unit 5 units Protease Inhibitor Cocktail 1 x 100µl 1 x 500µl Stop Solution 1 x 11ml 1 x 55ml Wild-type oligonucleotide (10 pmol/μL) 1 x 100µl 1 x 500µl
FunctionTranscriptional activator that binds to the consensus sequence 5'-AGATAG-3' and plays a key role in cardiac development (PubMed:24000169). Involved in bone morphogenetic protein (BMP)-mediated induction of cardiac-specific gene expression (By similarity). Binds to BMP response element (BMPRE) DNA sequences within cardiac activating regions (By similarity). Acts as a transcriptional activator of ANF in cooperation with NKX2-5 (By similarity). Promotes cardiac myocyte enlargement (PubMed:20081228). Required during testicular development (PubMed:21220346). May play a role in sphingolipid signaling by regulating the expression of sphingosine-1-phosphate degrading enzyme, spingosine-1-phosphate lyase (PubMed:15734735).
Involvement in diseaseAtrial septal defect 2
Ventricular septal defect 1
Tetralogy of Fallot
Atrioventricular septal defect 4
Testicular anomalies with or without congenital heart disease
GATA4 mutations can predispose to dilated cardiomyopathy (CMD), a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death.
Sequence similaritiesContains 2 GATA-type zinc fingers.
modificationsMethylation at Lys-300 attenuates transcriptional activity.
- Information by UniProt
- GATA 4
- GATA binding protein 4
ab207206 has not yet been referenced specifically in any publications.