Western blot analysis of extracts from various cell lines using Ras (G12V Mutant Specific) (D2H12) Rabbit mAb (upper), Ras (D2C1) Rabbit mAb #8955 (middle), and β-Actin (D6A8) #8457 (lower).
Western blot analysis of extracts from various cell lines using Ras (G12D Mutant Specific) (D8H7) Rabbit mAb (upper), Ras (D2C1) Rabbit mAb #8955 (middle), and β-Actin (D6A8) #8457 (lower).
Western blot analysis of extracts from various cell lines using Ras (D2C1) Rabbit mAb.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
|Ras (G12V Mutant Specific) (D2H12) Rabbit mAb 14412||20 µl||
|Ras (G12D Mutant Specific) (D8H7) Rabbit mAb 14429||20 µl||
|Ras (D2C1) Rabbit mAb 8955||20 µl||
||H Mk M R||21||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The Mutant Ras Antibody Sampler Kit provides an economical means of detecting common mutation in the Ras protein. The kit contains enough primary and secondary antibodies to perform at least two western blot experiments.
Each antibody in the Mutant Ras Antibody Sampler kit detects endogenous levels of its target protein. Ras (D2C1) Rabbit mAb recognizes endogenous levels of total Ras protein, specifically K-Ras and N-Ras. This antibody does not cross-react with H-Ras or R-Ras.
Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to G12V and G12D mutant sequences of human Ras protein. Ras (D2C1) Rabbit mAb was produced by immunizing rabbits with a recombinant protein specific to human K-Ras protein.
The 21 kDa guanine-nucleotide binding proteins (K-Ras, H-Ras, and N-Ras) cycle between active (GTP-bound) and inactive (GDP-bound) forms (1). Receptor tyrosine kinases and G protein-coupled receptors activate Ras, which then stimulates the Raf-MEK-MAPK pathway (2-4). GTPase-activating proteins (GAP) normally facilitate the inactivation of Ras. However, research studies have shown that in 30% of human tumors, point mutations in Ras prevent the GAP-mediated inhibition of this pathway (5). The most common oncogenic Ras mutation found in tumors is Gly12 to Asp12 (G12D), which prevents Ras inactivation, possibly by increasing the overall rigidity of the protein (5,6). Additional oncogenic mutations of Ras have been observed at varying frequencies at codons 12, 13, and 61. The Gly12 to Val12 (G12V) mutation has been detected in a number of different cancers, including colorectal and thyroid cancer (7,8).
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