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Mouse Monoclonal to H-Ras

H-RAS-03 (IgG1)

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Background

H-Ras is one of three ubiquitously isoforms of Ras GTPase that operate at the intracellular leaflet of the plasma membrane to regulate multiple signal transduction pathways, such as mitogen-activated protein kinase (MAPK) cascade. H Ras is anchored to the plasma membrane by farnesyl and two palmityl residues. GTP loading decreases H-Ras affinity for lipid rafts and allows the protein to target to nonraft microdomains, the primary sites of H-Ras signaling. Sos protein and other guanine nucleotide-exchange factors catalyze dissociation of GDP from Ras. Besides its roles in the plasma membrane, active H-Ras also diffuses through the cytoplasm on nanoparticles termed rasosomes, which is dependent on Ras palmitoylation.

Specificity

The antibody H-RAS-03 reacts with human H-Ras, an ubiquitously expressed 21 kDa intracellular protein. Although reactivity with other species has not been determined, it is probable as the epitope is highly conserved among animals.

Regulatory Status

Immunogen

Peptide corresponding to amino acids DIHQYREQIKRVKDSDDC of human H-Ras protein

Species Reactivity:

  • Human
  • Other not determined

Negative Species:

Applications:

  • Western Blotting
    Recommended dilution: 0.5 - 1 μg/ml
    Positive control:
    RAJI human Burkitt lymphoma cell line
    HeLa human cervix carcinoma cell line
    K567 human leukemia cell line
    Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with reducing Laemmli SDS-PAGE sample buffer. Boil for 5 min.
    Application note: Reducing conditions.
Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

General references

  • *Prior IA, Harding A, Yan J, Sluimer J, Parton RG, Hancock JF: GTP-dependent segregation of H-ras from lipid rafts is required for biological activity. Nat Cell Biol. 2001 Apr;3(4):368-75. [Abstract]
  • *Zheng Y, Quilliam LA: Activation of the Ras superfamily of small GTPases. Workshop on exchange factors. EMBO Rep. 2003 May;4(5):463-8. [Abstract] [Full Text]
  • *Rotblat B, Prior IA, Muncke C, Parton RG, Kloog Y, Henis YI, Hancock JF: Three separable domains regulate GTP-dependent association of H-ras with the plasma membrane. Mol Cell Biol. 2004 Aug;24(15):6799-810. [Abstract] [Full Text]
  • *Roy S, Plowman S, Rotblat B, Prior IA, Muncke C, Grainger S, Parton RG, Henis YI, Kloog Y, Hancock JF: Individual palmitoyl residues serve distinct roles in H-ras trafficking, microlocalization, and signaling. Mol Cell Biol. 2005 Aug;25(15):6722-33. [Abstract] [Full Text]
  • *Rotblat B, Yizhar O, Haklai R, Ashery U, Kloog Y: Ras and its signals diffuse through the cell on randomly moving nanoparticles. Cancer Res. 2006 Feb 15;66(4):1974-81. [Abstract] [Full Text]
  • For research use only. Not for drug, diagnostic or other use.

    Related Products

  • Mouse IgG1 Isotype Control
  • Example Data


    H-Ras-03
    Fig. 1. Western Blotting



    Fig. 1. Western Blotting analysis (reducing conditions) of H-Ras in whole cell lysate using anti-H-Ras (H-RAS-03).
    Lane 1: HeLa human cervix carcinoma cell line
    Lane 2: K567 human leukemia cell line
    Lane 3: RAJI human Burkitt lymphoma cell line


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