TIGAR is a p53-inducible enzyme that catalyzes the hydrolysis of fructose-2–6 bisphosphate (F-2–6-BP) to fructose-6-phosphate and inorganic phosphate. F-2–6-BP is a powerful activator of 6-phosphofructose-1 kinase, the rate limiting enzyme of glycolysis. By lowering the intracellular level of F-2–6-BP, TIGAR expression leads to increased glucose processing via the pentose phosphate pathway, the major cellular source for NADPH. Protein transduction using TAT fusion proteins represents an alternative methodology for introducing transcription factors and other intracellular proteins into primary, as well as transformed, cells. Recombinant Human TIGAR-TAT expressed in E. coli is a 31.6 kDa protein containing 283 amino acid residues, including the 269 residues of full-length TIGAR fused to a 14-residue C-terminal peptide containing the TAT transduction domain (GGGYGRKKRRQRRR).
Amino Acid Sequence
Pretreatment with TIGAR-TAT for 4 hrs, using a concentration range 0.1–5.0 µg/ml, protects U2OS cells from apoptosis induced by hydrogen peroxide.
Endotoxin level is <0.1 ng/μg of protein (<1EU/μg).
Centrifuge the vial prior to opening. Reconstitute in water to a concentration of 0.1–1.0 mg/ml. Do not vortex. For extended storage, it is recommended to further dilute in a buffer containing a carrier protein (example 0.1% BSA) and store in working aliquots at –20°C to –80°C.