Specificity
Clone 3H6 is a mouse monoclonal antibody that specifically detects human GTP-binding protein Rheb.
Application
Anti-Rheb, clone 3H6, Cat. No. MABC1605, is a mouse monoclonal antiboody that detects GTP-binding protein Rheb and has been tested for use in Western Blotting.
Western Blotting
Western Blotting Analysis: A representative lot detected Rheb in Western Blotting applications (Findlay, G.M., et. al. (2007). Biochem J. 403(1):13-20; Groenewoud, M.J., et. al. (2013). PLoS One. 8(11):e81649.
Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Rheb in MCF-7, JEG-3, and HT-29 cell lysates.
Features and Benefits
GST-tagged recombinant fragment corresponding to the first 167 amino acids of human GTP-binding protein Rheb.
Storage and Stability
Stable for 1 year at 2-8°C from date of receipt. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Analysis Note
Purified mouse monoclonal antibodyIgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Other Notes
GTP-binding protein Rheb (UniProt: Q15382; also known as Ras homolog enriched in brain) is encoded by the RHEB (also known as RHEB2) gene (Gene ID: 6009) in human. Rheb is a highly conserved small GTPase with low intrinsic GTPase activity. It is ubiquitous in its distribution, but higher levels of Rheb are reported in the brain, and in skeletal and cardiac muscles. It is synthesized with a short propeptide (aa 182-184), which is cleaved off to generate the active form. It is considered as a critical regulator of mTORC1 activity and is shown to bind to mTORC1 in a guanyl nucleotide-independent manner. It stimulates the phosphorylation of S6K1 and EIF4EBP1 through activation of mTORC1 signaling. Rheb is regulated in a negative manner by the GTPase-activating protein activity of a complex consisting of tuberous sclerosis complex 1 (TSC1) and TSC2. Akt or ERK1/2 are reported to phosphorylate and inactivate TSC that leads to the activation of Rheb and mTORC1. (Ref.: Findlay, GM et al. (2007) Biochem. J. 403(1); 13-20).
Evaluated by Western Blotting in lysate from DLD-1 cells.
Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Rheb in lysate from DLD-1 cells.
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