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|> 95% by SDS-PAGE & HPLC analyses
|< 0.1 ng/µg of protein (<1EU/µg)
|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.
|10mM Sodium Phosphate, pH 7.5
|Centrifuge the vial prior to opening. Reconstitute in water to a concentration of 0.1-1.0 mg/ml. Do not vortex. This solution can be stored at 2-8°C for up to 1 week. 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.
|Stability and Storage
|The lyophilized protein is stable at room temperature for 1 month and at 4°C for 3 months. Reconstituted working aliquots are stable for 1 week at 2°C to 8°C and for 3 months at -20°C to -80°C.
|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 36 kDa protein containing 284 amino-acid residues, including the 271 residues of full-length TIGAR fused to a 13-residue C-terminal peptide containing the TAT transduction domain (GGYGRKKRRQRRR).
|ARFALTVVRH GETRFNKEKI IQGQGVDEPL SETGFKQAAA AGIFLNNVKF THAFSSDLMR TKQTMHGILE RSKFCKDMTV KYDSRLRERK YGVVEGKALS ELRAMAKAAR EECPVFTPPG GETLDQVKMR GIDFFEFLCQ LILKEADQKE QFSQGSPSNC LETSLAEIFP LGKNHSSKVN SDSGIPGLAA SVLVVSHGAY MRSLFDYFLT DLKCSLPATL SRSELMSVTP NTGMSLFIIN FEEGREVKPT VQCICMNLQD HLNGLTETRG GGYGRKKRRQ RRR
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