Rheb Antibody (JG37-12)

Western Blot: Rheb Antibody (JG37-12) [NBP2-75646] - Analysis of Rheb on mouse placenta tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry/Immunofluorescence: Rheb Antibody (JG37-12) [NBP2-75646] - Staining RHEB in SH-SY-5Y cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Immunohistochemistry-Paraffin: Rheb Antibody (JG37-12) [NBP2-75646] - Analysis of paraffin-embedded human fetal skeletal muscle tissue using anti-RHEB antibody. Counter stained with hematoxylin.

Flow Cytometry: Rheb Antibody (JG37-12) [NBP2-75646] - Analysis of A431 cells with RHEB antibody at 1/100 dilution (yellow) compared with an unlabelled control (cells without incubation with primary antibody; purple).Alexa Fluor 488-conjugated goat anti-rabbit IgG was used as the secondary antibody.

Immunocytochemistry/Immunofluorescence: Rheb Antibody (JG37-12) [NBP2-75646] - Staining RHEB in A431 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Immunohistochemistry-Paraffin: Rheb Antibody (JG37-12) [NBP2-75646] - Analysis of paraffin-embedded human liver tissue using anti-RHEB antibody. Counter stained with hematoxylin.

Immunohistochemistry-Paraffin: Rheb Antibody (JG37-12) [NBP2-75646] - Analysis of paraffin-embedded human colon tissue using anti-RHEB antibody. Counter stained with hematoxylin.

Knockdown Validated: Rheb Antibody (JG37-12) [NBP2-75646] - Analysis of RHEB with anti-RHEB antibody [JG37-12] at 1:1,000 dilution. Lane 1: Wild-type Raw264.7 whole cell lysate. Lane 2: RHEB knockdown Raw264.7 whole cell lysate. NBP2-75646 was shown to specifically react with RHEB in wild-type Raw264.7 cells. Weakened band was observed when RHEB knockout samples were tested. Wild-type and RHEB knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-RHEB antibody ( 1/1,000) and Anti-GAPDH antibody ( 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature.

Product Discontinued

Rheb Antibody (JG37-12) Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Recombinant protein within Human Rheb aa 21-108 / 184. (SwissProt: Q15382 Human; SwissProt: Q921J2 Mouse)
• Localization: Cytoplasm. Golgi apparatus. Endoplasmic reticulum.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: RHEB
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:50-1:100
  • Immunocytochemistry/Immunofluorescence 1:50-1:200
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:50-1:200
  • Knockdown Validated
  • Western Blot 1:500-1:2000
• Theoretical MW: 20 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: TBS (pH7.4), 0.05% BSA, 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for Rheb Antibody (JG37-12)

• GTP-binding protein Rheb
• MGC111559
• Ras homolog enriched in brainRHEB2Ras homolog enriched in brain 2
• Rheb
• Rheb2

Background

Rheb (Ras homolog enriched in brain) is an evolutionarily conserved member of the Ras family of small GTP binding proteins originally found to be rapidly induced by synaptic activity in the hippocampus following seizure. While it is expressed at relatively high levels in the brain, Rheb is widely expressed in other tissues and may be induced by growth factor stimulation. Similar to other family members, Rheb triggers activation of the Raf-MEK-MAPK pathway. Biochemical and genetic studies demon-strate that Rheb has an important role in regulating the insulin/Target of rapamycin (TOR) signaling pathway. TOR is a serine/threonine protein kinase that acts as a sensor for ATP and amino acids, balancing the availability of nutrients with protein translation and cell growth. A dimeric protein complex termed TSC1/TSC2 indirectly inhibits TOR activity by inhibiting Rheb via the GAP activity of TSC2.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: RHEB