RHEB Antibody (YA094)
(Synonyms: RHEB2, RHEB, GTP-binding protein Rheb, Ras homolog enriched in brain)Based on 1 Customer Validation
RHEB Antibody (YA094) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to RHEB.
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Host:
Rabbit
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Isotype:
IgG
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Application:
WB, ICC/IF, IHC-P, FC
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Reactivity :
Human, Mouse
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Formulation:
Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.
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Conjugation:
Non-conjugated
Applications
| Application |
WB
WB: Western Blot
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ICC/IF
ICC/IF: Immunocytochemistry/
Immunofluorescence |
IHC-P
IHC-P: Immunohistochemistry-Paraffin
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FC
FC: Flow Cytometry
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| Dilution Ratio | 1:500-1:2000 | 1:50-1:200 | 1:50-1:200 | 1:50-1:100 |
Product Details
RHEB Antibody (YA094) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to RHEB.
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Host Rabbit
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Clonality Recombinant,Monoclonal
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Species ReactivityHuman, Mouse
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Observed Molecular WeightObserved band size: 27 kDaNote: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
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Calculated Molecular Weight Predicted band size: 20 kDa;
Synthetic peptide corresponding to Human RHEB.AA range:21-108.
Endogenous
Protein A affinity purified.
Non-conjugated
Unmodified
IgG
Product Properties
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Appearance
Solution
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Formulation
Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.
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Concentration
Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration
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Storage & Stability
Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.
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Shipping
Shipping with blue ice.
Verification Images
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Western blot analysis of extracts from M-brain using RHEB antibody. Proteins were transferred to a PVDF membrane and blocked with 5% nonfat powdered milk in PBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (GAPDH, 1/3000) was diluted with 5% nonfat powdered milk in PBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/8,000) was incubated for 45min at room temperature. -
Immunohistochemical analysis of paraffin-embedded mouse colon using RHEB antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80310, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody (HY-P83652). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded mouse liver using RHEB antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80310, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody (HY-P83652). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded mouse stomach using RHEB antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80310, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody (HY-P83652). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded mouse heart using RHEB antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80310, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody (HY-P83652). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded mouse brain using RHEB antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80310, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody (HY-P83652). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded mouse prostate using RHEB antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80310, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody (HY-P83652). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of 1X106 HeLa cells labeling RHEB Antibody (HY-P80310, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/100 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).
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Immunocytochemistry analysis of HeLa cells labeling RHEB with RHEB Antibody (HY-P80310) at 1/100 dilution . Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with RHEB Antibody (HY-P80310) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L(HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
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Immunocytochemistry analysis of A431 cells labeling RHEB with RHEB Antibody (HY-P80310) at 1/100 dilution . Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with RHEB Antibody (HY-P80310) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L(HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
Background
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Function
Small GTPase that acts as an allosteric activator of the canonical mTORC1 complex, an evolutionarily conserved central nutrient sensor that stimulates anabolic reactions and macromolecule biosynthesis to promote cellular biomass generation and growth (PubMed:12172553, PubMed:12271141, PubMed:12842888, PubMed:12869586, PubMed:12906785, PubMed:15340059, PubMed:15854902, PubMed:16098514, PubMed:20381137, PubMed:22819219, PubMed:24529379, PubMed:29416044, PubMed:32470140, PubMed:33157014, PubMed:25816988). In response to nutrients, growth factors or amino acids, specifically activates the protein kinase activity of MTOR, the catalytic component of the mTORC1 complex: acts by causing a conformational change that allows the alignment of residues in the active site of MTOR, thereby enhancing the phosphorylation of ribosomal protein S6 kinase (RPS6KB1 and RPS6KB2) and EIF4EBP1 (4E-BP1) (PubMed:29236692, PubMed:33157014). RHEB is also required for localization of the TSC-TBC complex to lysosomal membranes (PubMed:24529379). In response to starvation, RHEB is inactivated by the TSC-TBC complex, preventing activation of mTORC1 (PubMed:24529379, PubMed:33157014). Has low intrinsic GTPase activity (PubMed:15340059)
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Subcellular Localization
Endomembrane system; Lipid-anchor; Cytoplasmic side; Lysosome membrane; Lipid-anchor; Cytoplasmic side; Golgi apparatus membrane; Lipid-anchor; Cytoplasmic side; Endoplasmic reticulum membrane; Lipid-anchor; Cytoplasmic side; Cytoplasm, cytosol
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Expression
Tissue_specificity:Ubiquitous (PubMed:8543055) . Highest levels observed in skeletal and cardiac muscle (PubMed:8543055) -
Subunit
Associates with the mTORC1 complex (MTOR, MLST8 and RPTOR) in a guanyl nucleotide-independent manner (PubMed:15854902, PubMed:16098514, PubMed:24529379). Interacts with TSC2 (PubMed:15854902, PubMed:22819219, PubMed:24529379, PubMed:25816988). Interacts with MCRS1; the interaction maintains RHEB at the lysosome in its active GTP-bound form and prevents its interaction with the mTORC1 complex inhibitor TSC2, ensuring activation of the mTORC1 complex by RHEB (PubMed:25816988). Interacts (when prenylated) with PDE6D; this promotes release from membranes (PubMed:22002721)
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SwissProt ID
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Synonyms
RHEB2, RHEB, GTP-binding protein Rheb, Ras homolog enriched in brain
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Research Field
Signal Transduction
Documentation
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Data Sheet (235 KB)
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SDS (313 KB)
- English - EN (313 KB)
- Français - FR (313 KB)
- Deutsch - DE (313 KB)
- Norwegian - NO (313 KB)
- Español - ES (313 KB)
- Swedish - SV (313 KB)
- Italian - IT (313 KB)
- Korean - KR (313 KB)
- Portuguese - PT (313 KB)
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User Guide for Antibodies (1077 KB)