Type
Recombinant
Description
TIGARisap53-inducibleenzymethatcatalyzesthehydrolysisoffructose-2–6bisphosphate(F-2–6-BP)tofructose-6-phosphateandinorganicphosphate.F-2–6-BPisapowerfulactivatorof6-phosphofructose-1 kinase,theratelimitingenzymeofglycolysis.ByloweringtheintracellularlevelofF-2–6-BP,TIGARexpressionleadstoincreasedglucoseprocessingviathepentosephosphatepathway,themajorcellularsourceforNADPH.ProteintransductionusingTATfusionproteinsrepresentsanalternativemethodologyforintroducingtranscriptionfactorsandotherintracellularproteinsintoprimary,aswellastransformed,cells.RecombinantHumanTIGAR-TATexpressedinE.coliisa31.6 kDaproteincontaining283 aminoacidresidues,includingthe269 residuesoffull-lengthTIGARfusedtoa14-residueC-terminalpeptidecontainingtheTATtransductiondomain(GGGYGRKKRRQRRR).
AminoAcidSequence
ARFALTVVRHGETRFNKEKIIQGQGVDEPLSETGFKQAAAAGIFLNNVKFTHAFSSDLMRTKQTMHGILERSKFCKDMTVKYDSRLRERKYGVVEGKALSELRAMAKAAREECPVFTPPGGETLDQVKMRGIDFFEFLCQLILKEADQKEQFSQGSPSNCLETSLAEIFPLGKNHSSKVNSDSGIPGLAASVLVVSHGAYMRSLFDYFLTDLKCSLPATLSRSELMSVTPNTGMSLFIINFEEGREVKPTVQCICMNLQDHLNGLTETRGGGYGRKKRRQRRR
Source
E.coli
Purity
95%
BiologicalActivity
PretreatmentwithTIGAR-TATfor4 hrs,usingaconcentrationrange0.1–5.0 µg/ml,protectsU2OScellsfromapoptosisinducedbyhydrogenperoxide.
Endotoxin
Endotoxinlevelis<0.1 ng/μgofprotein(<1EU/μg).
Reconstitution
Centrifugethevialpriortoopening.Reconstituteinwatertoaconcentrationof0.1–1.0 mg/ml.Donotvortex.Forextendedstorage,itisrecommendedtofurtherdiluteinabuffercontainingacarrierprotein(example0.1%BSA)andstoreinworkingaliquotsat –20°Cto –80°C.
Storage/Expiration
–20°C