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IRDye 700 HIF-1 Consensus Oligonucleotide

5′ -- AGC TTG CCC TAC GTG CTG TCT CAG A -- 3′

3′ -- TCG AAC GGG ATG CAC GAC AGA GTC T -- 5′

Underlined nucleotides are the binding site. IRDye 700 oligonucleotides are supplied as 25 µL of 50 nM (or 50 fmol/µL) double-stranded DNA.

HIF-1 is a transcription factor that regulates celluar responses to hypoxia, which have been heavily implicated in cancer biology and various other pathophysiologies. For this reason, it is a common oligonucleotide used in EMSA assays in basic discovery and therapeutics research.

After the addition of the DNA to the protein-buffer mix, reactions are incubated to allow protein binding to DNA. A typical incubation condition is 20-30 minutes at room temperature.

Since IRDye infrared dyes are somewhat sensitive to light, it is best to keep binding reactions in the dark during incubation periods (e.g., put the tubes into a drawer or simply cover the rack containing tubes with aluminum foil). After the incubation period, 10X Orange Loading Dye (P/N 927-10100) is added to the binding reaction for electrophoresis.