GATA1 Transcription Factor Activity Assay

This Assay Genie kit completes the full workflow in approximately 4.5 hours from lysate loading to readable plate. The microplate is pre-coated with streptavidin-bound biotinylated double-stranded oligonucleotides, so the WGATAA capture sequence is immobilized directly onto each well. A lyophilized Nuclear Lysate Positive Control is included and is reconstituted in 110 µL of 1x Binding Buffer; running it alongside the optional Wild-Type and Mutant Consensus Oligonucleotide competitor controls confirms that any signal reflects sequence-specific binding rather than off-target interactions.

The supplied GATA1 primary antibody is provided at 100x concentration and is paired with an HRP-conjugated anti-rabbit IgG secondary for TMB-based detection; color development proceeds at room temperature on an orbital shaker and is terminated with stop solution, producing the expected blue-to-yellow shift. For upstream lysis, the protocol recommends supplementing the provided cytoplasmic and nuclear extraction buffers with freshly prepared PMSF and a protease inhibitor cocktail, then clarifying nuclear extracts by centrifugation at 3000 x g. Aliquot cleared nuclear extracts and store frozen at -80 °C, avoiding repeated freeze-thaw cycles. Unused microplate strips should be resealed and refrigerated at 4 °C, where the complete kit remains stable for up to 6 months from receipt.