Recombinant Anti-RHEB antibody [EPR2971] (ab92313)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2971] to RHEB
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-RHEB antibody [EPR2971]
See all RHEB primary antibodies -
Description
Rabbit monoclonal [EPR2971] to RHEB -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human RHEB aa 50-150. The exact sequence is proprietary.
(Peptide available asab194764) -
Positive control
- WB: Raji and SH-SY5Y lysates. IHC-P: human glioma and brain tissue. ICC/IF: Raji cells. Flow Cyt (intra): Raji cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2971 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab92313 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/40.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 - 1/1000 |
|
WB |
1/2000 - 1/5000. Predicted molecular weight: 20 kDa.Can be blocked with Human RHEB peptide (ab194764).
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IHC-P | (2) |
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Heat mediated antigen retrieval and use of an HRP/AP polymerized secondary antibody recommended. See IHC antigen retrieval protocols For unpurified use at 1/100 - 1/250 |
ICC/IF |
1/100 - 1/250.
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Notes |
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Flow Cyt (Intra)
1/40. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 - 1/1000 |
WB
1/2000 - 1/5000. Predicted molecular weight: 20 kDa.Can be blocked with Human RHEB peptide (ab194764). |
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Heat mediated antigen retrieval and use of an HRP/AP polymerized secondary antibody recommended. See IHC antigen retrieval protocols For unpurified use at 1/100 - 1/250 |
ICC/IF
1/100 - 1/250. |
Target
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Function
Stimulates the phosphorylation of S6K1 and EIF4EBP1 through activation of mTORC1 signaling. Activates the protein kinase activity of mTORC1. Has low intrinsic GTPase activity. -
Tissue specificity
Ubiquitous. Highest levels observed in skeletal and cardiac muscle. -
Sequence similarities
Belongs to the small GTPase superfamily. Rheb family. -
Post-translational
modificationsFarnesylation is important for efficiently activating mTORC1-mediated signaling. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 6009 Human
- Omim: 601293 Human
- SwissProt: Q15382 Human
- Unigene: 283521 Human
- Unigene: 568850 Human
- Unigene: 647068 Human
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Alternative names
- Ras homolog enriched in brain 2, formerly antibody
- GTP binding protein Rheb antibody
- GTP-binding protein Rheb antibody
see all
Images
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Anti-RHEB antibody [EPR2971] (ab92313) at 1/2000 dilution + Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 20 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue sections labeling RHEB with purified ab92313 at 1:500 dilution (0.73 µg/ml). Heat mediated antigen retrieval was performed using Bond ™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Intracellular Flow Cytometry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling RHEB with purified ab92313 at 1/40 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry/ Immunofluorescence analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling RHEB with purified ab92313 at 1:250 dilution (1.46 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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All lanes : Anti-RHEB antibody [EPR2971] (ab92313) at 1/2000 dilution
Lane 1 : Raji cell lsyate
Lane 2 : SH-SY5Y cell lsyate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 20 kDa -
Overlay histogram showing Raji cells stained with unpurifiedab92313 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92313, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Raji cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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Unpurified ab92313, at 1/100 dilution, staining RHEB in paraffin-embedded Human brain tissue, by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (8)
ab92313 has been referenced in 8 publications.
- Chen R et al. circRNA MYLK Accelerates Cervical Cancer via Up-Regulation of RHEB and Activation of mTOR Signaling. Cancer Manag Res 12:3611-3621 (2020). PubMed: 32547198
- Pang K et al. Resveratrol inhibits hypertrophic scars formation by activating autophagy via the miR-4654/Rheb axis. Mol Med Rep 22:3440-3452 (2020). PubMed: 32945452
- Liu F et al. Overexpression of RHEB is associated with metastasis and poor prognosis in hepatocellular carcinoma. Oncol Lett 15:3838-3845 (2018). PubMed: 29467900
- Zhou L et al. miR-185 Inhibits Fibrogenic Activation of Hepatic Stellate Cells and Prevents Liver Fibrosis. Mol Ther Nucleic Acids 10:91-102 (2018). PubMed: 29499960
- Shen Y et al. MicroRNA-194 overexpression protects against hypoxia/reperfusion-induced HK-2 cell injury through direct targeting Rheb. J Cell Biochem N/A:N/A (2018). PubMed: 30485514
- Zhou L et al. MicroRNA-185 induces potent autophagy via AKT signaling in hepatocellular carcinoma. Tumour Biol 39:1010428317694313 (2017). PubMed: 28240051
- Song Z et al. Resistance exercise initiates mechanistic target of rapamycin (mTOR) translocation and protein complex co-localisation in human skeletal muscle. Sci Rep 7:5028 (2017). PubMed: 28694500
- Jiang L et al. Rheb/mTORC1 Signaling Promotes Kidney Fibroblast Activation and Fibrosis. J Am Soc Nephrol 24:1114-26 (2013). IHC-P ; Human . PubMed: 23661807