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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.
More>>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. Less<<
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Übersicht
Replacement Information
Description
Catalogue Number
MABC1605-100UG
Description
Anti-Rheb Antibody, clone 3H6
Alternate Names
GTP-binding protein Rheb
Ras homolog enriched in brain
Background Information
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).
References
Product Information
Format
Purified
Presentation
Purified mouse monoclonal antibodyIgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Applications
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.
Key Applications
Western Blotting
Application Notes
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.
Biological Information
Immunogen
GST-tagged recombinant fragment corresponding to the first 167 amino acids of human GTP-binding protein Rheb.
Clone
3H6
Concentration
Please refer to lot specific datasheet.
Host
Mouse
Specificity
Clone 3H6 is a mouse monoclonal antibody that specifically detects human GTP-binding protein Rheb.
~20 kDa observed; 20.50 kDa calculated. Uncharacterized bands may be observed in some lysate(s).
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
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.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
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.
The Ras-like GTPase Rheb has been identified as a crucial activator of mTORC1. Activation most likely requires a direct interaction between Rheb and mTOR, but the exact mechanism remains unclear. Using a panel of Rheb-deficient mouse embryonic fibroblasts (MEFs), we show that Rheb is indeed essential for the rapid increase of mTORC1 activity following stimulation with insulin or amino acids. However, mTORC1 activity is less severely reduced in Rheb-deficient MEFs in the continuous presence of serum or upon stimulation with serum. This remaining mTORC1 activity is blocked by depleting the cells for amino acids or imposing energy stress. In addition, MEK inhibitors and the RSK-inhibitor BI-D1870 interfere in mTORC1 activity, suggesting that RSK acts as a bypass for Rheb in activating mTORC1. Finally, we show that this rapamycin-sensitive, Rheb-independent mTORC1 activity is important for cell cycle progression. In conclusion, whereas rapid adaptation in mTORC1 activity requires Rheb, a second Rheb-independent activation mechanism exists that contributes to cell cycle progression.
A MAP4 kinase related to Ste20 is a nutrient-sensitive regulator of mTOR signalling. Findlay, GM; Yan, L; Procter, J; Mieulet, V; Lamb, RF Biochem J
403
13-20
2007
The mTOR (mammalian target of rapamycin) signalling pathway is a key regulator of cell growth and is controlled by growth factors and nutrients such as amino acids. Although signalling pathways from growth factor receptors to mTOR have been elucidated, the pathways mediating signalling by nutrients are poorly characterized. Through a screen for protein kinases active in the mTOR signalling pathway in Drosophila we have identified a Ste20 family member (MAP4K3) that is required for maximal S6K (S6 kinase)/4E-BP1 [eIF4E (eukaryotic initiation factor 4E)-binding protein 1] phosphorylation and regulates cell growth. Importantly, MAP4K3 activity is regulated by amino acids, but not the growth factor insulin and is not regulated by the mTORC1 inhibitor rapamycin. Our results therefore suggest a model whereby nutrients signal to mTORC1 via activation of MAP4K3.
