Anti-Ras Antibody, clone RAS10, Alexa Fluor^™ 555 Conjugate

Ras proteins are small GTPases controlling cellular pathways that signal for growth, proliferation, and differentiation. Mammalian genomes harbor three Ras genes (HRAS, NRAS, and KRAS) that encode four GTPases, two of which (K-Ras4A and K-Ras4B) are transcribed from the KRAS gene via two alternative fourth exons. Ras proteins share nearly identical sequence homology in sequence encoded by exons 1-3 (a.a. 1-165), while diverge in their C-terminal 24-a.a. hypervariable region (HVR) encoded by exon 4. The first 165 amino acids form the catalytic G domain that mediates GDP/GTP binding and association with effectors, exchange factors, and GTPase-activating proteins (GAPs), while the C-terminal HVR is needed for Ras membrane localization, a process necessary for Ras function. In addition to the removal of initiator methionine (Met1), Ras proteins are posttranslationally modified by farnesylation of Cys186, followed by proteolytical cleavage their C-terminal end three amino acids (a.a. 187-189) and carboxyl methylation of the newly formed C-terminal prenylcysteine. H-Ras, N-Ras, and K-Ras4A are further palmitoylated on cysteine residue(s) near the C-terminus (Cys181/Cys184 in H-Ras, Cys181 in N- and K-Ras), while K-Ras4B lacks a palmitoylation site. Palmitoylation is required for targeting H- and N-Ras to the plasma membrane (PM), while K-Ras4A and K-Ras4B contain a series of lysines that constitute a polybasic region (PBR) capable of interacting with the inner leaflet of the PM through an electrostatic interaction.

Ras 10 reacts with normal p21, activated p21, and p21 from each of the Harvey, Kirsten and N-ras families. Ras 10 reacts with recombinant p21 as well as p21 derived from human and rodent cells (Hamer, P.J., et al. (1990). Hybridoma.9(6):573-587).

Recombinant human H-Ras p21 (Hamer, P.J., et al. (1990). Hybridoma.9(6):573-587).

Detect Ras using this mouse monoclonal Anti-Ras Antibody, clone RAS10, Alexa Fluor^™ 555 Conjugate, Cat. No. 05-516-AF555, validated for use in Flow Cytometry aned Immunocytochemistry.

Flow Cytometry Analysis: 0.2 µL from a representative lot detected Ras immunoreactivity in one million A431 cells.

Clone Ras10 is also available as unconjugated, as well as in FITC- and Alexa Fluor^™ 647-conjugated forms (Cat. Nos. 05-516, FCMAB148F, 05-516-AF647) for ELISA, flow cytometry, immunocytochemistry, immunohistochemistry, immunoprecipitation, and Western blotting applications.

Research Category
Signaling

Evaluated by Immunocytochemistry in A431 cells.

Immunocytochemistry Analysis: A 1:100 dilution of this antibody detected Ras immunoreactivity in A431 cells.

~21 kDa observed. 21.30 kDa (human/mouse/rat H-Ras4A, p21 pro-form), 20.87 kDa (human/mouse/rat mature H-Ras4A, p21), 21.66 kDa (human/mouse/rat K-Ras4A pro-form), 21.17 kDa (human/mouse/rat mature K-Ras4A), 21.23/21.20/21.24 kDa (human/mouse/rat N-Ras pro-form), 20.90/20.90/20.91 kDa (human/mouse/rat mature N-Ras) calculated.

Protein G purified.

Purified mouse IgG2a Alexa Fluor^™ 555 conjugate in buffer containing PBS with 1.5% BSA and 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Concentration: Please refer to lot specific datasheet.

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