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The GATA family consists of six members, GATA 1-6, that bind to the (A/T)GATA(A/G) nucleotide motif. GATA family members are grouped into two subfamilies based on structural and expression comparisons. GATA-1, -2 and -3 are expressed predominantly in hematopoietic cells, and regulate differentiation and gene expression in T-lymphocytes, erythroids and megakaryocytes. GATA-4, -5 and -6 are found mainly in the heart and gut, and are involved in regulation of cardiogenesis and gut development. The TransAM GATA Family Kit measures GATA-1, -2 and –3, while TransAM GATA-4 is specific for studying GATA-4.
Figure 1: Monitoring GATA family member activation using the TransAM GATA Family Kit.
GATA-1, -2 and -3 activation were assayed using the TransAM GATA Family Kit. 1:1000 dilutions of each antibody in the kit were tested using 5 µg/well of nuclear extract prepared from an unstimulated cell line: GATA-1 and -2 were tested with K-562 and GATA-3 with Jurkat. Assays were performed in the absence or presence of 20 pmol of competitor oligonucleotide that contains either a wild-type or mutated GATA consensus binding site. 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.
