Human Ras Antibody (342404)

Detection of Human Ras by Western Blot.

Western blot shows lysates of K562 human chronic myelo-genous leukemia cell line and Ramos human Burkitt's lymphoma cell line. PVDF membrane was probed with 5 µg/mL Rat Anti-Human Ras Monoclonal Antibody (Catalog # MAB3429) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). For additional refer-ence, recombinant human H-Ras, N Ras, K-Ras2A and 2B, R-Ras2 and 3, Rap2A and 2B, RalA and B, and Rheb (5 ng/lane) were included. A specific band for Ras was detected at approximately 21 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.

Detection of Ras by Immunocytochemistry/ Immunofluorescence

The level of TRPC3 in fibroblasts affects basal mitochondrial Ca2+ load.a The gallery (left) shows (from top to bottom) the transmitted light image of a single fibroblast, confocal image of GEM-GECO1mito fluorescence captured at wavelengths 470–500 nm, at wavelengths >520 nm and their overlay. The gallery (middle) compares confocal images (overlay of 2 channels: BP 470–500 nm plus LP > 520 nm) of GEM-GECO1mito fluorescence from the fibroblasts transduced with shCTL and shTRPC3, as indicated. The bar diagram plot (right) compares mean resting [Ca2+]mito (estimated as F470–500/F>520) in the fibroblasts transduced with shCTL (n = 96) and shTRPC3 (n = 96). ***P < 0.001 relative to shCTL (Student’s t test, two-sided). b Elevation of mitochondrial Ca2+ load upon hRAS overexpression is markedly prevented by simultaneous overexpression of TRPC3. The gallery (left) compares confocal images (overlay of 2 channels: BP 470–500 nm plus LP > 520 nm) of GEM-GECO1mito fluorescence from control MRC5, MRC5 with hRAS-induced senescence (hRAS) and MRC5 overexpressing hRAS and TRPC3 simultaneously (hRAS + TRPC3) or TRPC3 only (TRPC3). Images of three representative fields of view per condition are shown. The bar diagram plot (right) compares mean resting [Ca2+]mito (estimated as F470–500/F>520) in control MRC5 (Control: n = 390) with that in MRC5 with hRAS-induced senescence (+hRAS: n = 232) and MRC5 overexpressing hRAS and TRPC3 simultaneously (+hRAS + TRPC3: n = 266) or TRPC3 only (+TRPC3: n = 251). ***P < 0.001 relative to control, unless stated otherwise. One-way ANOVA, Tukey’s multiple comparisons test. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41467-022-28597-x), licensed under a CC-BY license. Not internally tested by R&D Systems.