mTOR/MLST8/RPTOR Complex Recombinant

Catalog #

102073

$525 *

Size: 10 µg

Qty

Bulk/Custom

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Description

Complex of mTOR (mammalian target of rapamycin), encompassing amino acids 1362-2549, full length MLST8 (target of rapamycin complex subunit LST8) and full length RPTOR (regulatory-associated protein of mTOR). The mTOR construct contains an N-terminal FLAG-tag, while MLST8 and RPTOR include an N-terminal Strep-tag. The proteins were expressed, and affinity purified as a complex.

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Synonyms

mTOR, mechanistic target of rapamycin, FRAP, FKBP-Rapamycin Associated Protein, Mammalian Target Of Rapamycin, raptor, regulatory associated protein of mTOR, Mip1, MLST8, Mammalian Lethal With SEC13 Protein 8, GBL, Gable, POP3, LST8

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

mTOR Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

mTOR (FLAG-1362-2549(end)) / MLST8 (Strep-2-326(end)) / RPTOR (Strep-2-1335(end))

Host Species/Expression System

Sf9

Purity

≥80%

Format

Aqueous buffer solution.

Formulation

80 mM Tris-HCl, pH 8.0, 120 mM NaCl, 0.8 mM EDTA, 2 mM desthiobiotin, and 20% Glycerol

mTOR: 137 kDa; MLST8: 37 kDa; RPTOR: 150 kDa

Amino Acids

mTOR: 1362-2549(end); MLST8: 2-326(end); RPTOR: 2-1335(end)

Genbank #

mTOR: NM_004958; MLST8: NM_022372; RPTOR: NM_020761

UniProt #

mTOR: NM_004958; MLST8: NM_022372; Raptor: NM_020761

Tag(s)

N-Terminal FLAG-Tag, N-Terminal Strep-Tag

Background

mTOR (mammalian target of rapamycin) is a serine/threonine kinase protein of the PIK3 (phosphatidylinositol 3- kinase)-related family. It can form two different complexes, which differ in the accessory protein present and have different biological functions. mTORC1 (mTOR complex 1) includes the protein RPTOR (regulatory-associated protein of mTOR), while mTORC2 is composed of RICTOR (RPTOR independent companion of mTOR complex 2). MLST8 (target of rapamycin complex subunit LST8) is common to both complexes and seems to stabilize the active site of mTOR and participate in its activation. mTORC1 is activated in response to growth factors and responds to activation of the PI3K-AKT (protein kinase B) pathway. It is involved in cell growth via phosphorylation of proteins like 4E-BP1 (eIF-4E-binding protein). Dysfunction of mTORC1 can result in cancer, and strategies targeting mTORC1 may result in major advances in cancer therapy.

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

Activity was measured using the ADP-Glo™ Kinase Assay (Promega #V6930). The kinase reaction included 100 µM ATP, 0.1 mg/ml 4EBP1, and 5 mM MnCl₂ in kinase buffer (40 mM Tris HCl, pH 7.4, 20 mM MgCl₂, 0.1 mg/ml BSA, and 2 mM DTT) (25 µl total volume per reaction). Reaction was incubated for 45 minutes at 30°C.

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles

Scientific Category

Cancer Therapy Target

Regulatory Status

Research Use Only