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Encompassing Amino Acids: mTOR 1362-end, MLST8 2-326 (end), Raptor 2-1335 (end)
Application: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Enzyme reaction is conducted in a buffer containing 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 200 µM ATP, 0.01% Brij-35 and 2 µM substrate (Invitrogen Zlyte Ser/Thr peptide 11) at room temperature for 1 hour.
Description: Complex of human mTOR, also known as FRAP1 (GenBank Accession No. NM_004958), a.a. 1362-end, with N-terminal FLAG-tag, MW = 137 kDa, human MLST8 (GenBank Accession No. NM_022372), a.a. 2-326 (end) with N-terminal His-tag, MW = 37 kDa, and human Raptor (GenBank Accession No. NM_020761), a.a. 2-1335 (end) with N-terminal Histag, MW=150 kDa, co-expressed in a Baculovirus-infected Sf9 cell expression system.
Format: Aqueous buffer solution
Formulation: 40 mM Tris, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 80 ng/µL FLAG peptide, and 20% glycerol.
Genbank: NM_004958 (mTOR), NM_022372 (MLST8), NM_020761 (Raptor)
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal FLAG-tag (mTOR), N-terminal His-tag (MLST8), N-terminal His-tag (Raptor)
Uniprot: P42345 (mTOR), Q9BVC4 (MLST8), Q8N122 (Raptor)
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Toral-Barza et al., (2005) Biochem. Biophys. Res. Commun. 24, 332(1):304-10.
2. Rubio-Viqueira and Hidalgo, M. (2006) Adv. Exp. Med. Biol. 587:309-327.Application Reference(s):
1. mTOR signaling regulates nucleolar targeting of the SUMO-specific isopeptidase SENP3 (2014)
2.The mechanistic target for rapamycin pathway is related to the phosphorylation score for estrogen receptor-α in human breast tumors in vivo (2014)
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