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Western blot analysis of mouse SYF cells transformed with c-Src then left untreated (lanes 1, 3, & 5) or treated with pervanadate (1 mM) for 30 minutes (lanes 2, 4, & 6). The blot was probed with anti-c-Src (Tyr-215) (lanes 1 & 2), anti-c-Src (N-terminal region) (lanes 3 & 4), and anti-c-Src (Tyr-530) (lanes 5 & 6).
Western blot image of mouse F9 stem cells treated with with calyculin A (100 nM, 30 min.) (lanes 1-4) then Sox2 was dephosphorylated with lambda phosphatase (lanes 2 & 4). The blot was probed with mouse monoclonal Sox2 (lanes 1 & 2) and rabbit polyclonal anti-Sox2 (Thr-118) phospho-specific antibody (lanes 3 & 4).Immunocytochemical labeling of Sox2 in aldehyde fixed and NP-40 permeabilized human NCI-H446 lung carcinoma cells. The cells were labeled with mouse monoclonal anti-Sox2 (SM5511). The antibody was detected using goat anti-mouse DyLight® 594.
Western blot of nSMase3a and nSMase3b GFP transfectants (A) and nSMase3 siRNA knockdown in C2C12 myotubes (B). Both blots were probed with anti-nSMase3 (C-terminal region) rabbit polyclonal antibody (SP0281). (Image provided by Dr. Jennifer Moylan, Dept. Physiology, University of Kentucky).Immunocytochemical labeling of nSMase3 in aldehyde-fixed and NP-40-permeabilized mouse C2C12 cells. The cells were labeled with rabbit polyclonal anti-nSMase3 (SP0281) antibody. The antibody was detected using appropriate secondary antibody conjugated to DyLight® 594.
Western blot of adult mouse brain. The blots were probed with anti-nSMase2 (C-terminal region) rabbit polyclonal antibody at 1:2000 (lane 1), 1:1000 (lane 2), or 1:500 in the absence (lane 3) or presence of nSMase2 blocking peptide (SX4065) (lane 4).Immunocytochemical labeling of nSMase2 in aldehyde-fixed and NP-40-permeabilized differentiated PC12 cells. The cells were labeled with rabbit polyclonal anti-nSMase2 (SP4061) antibody in the absence (Left) or presence (Right) of blocking peptide (SX4065). The antibody was detected using appropriate secondary antibody conjugated to DyLight® 594. The cells were counterstained with Sytox green to label nuclei.
Western blot analysis of SMAD7 in human HepG2 cells. The blot was probed with rabbit polyclonal SMAD7 antibody (SP4901) at 1:250.
Western blot analysis of adult mouse brain. The blot was probed with anti-SHP2 (N-terminal) antibody at 1:250 (lane 1), 1:500 (lane 2), 1:1000 (lane 3), and 1:2000 (lane 4).
Western blot analysis of human Jurkat cells treated with pervanadate (1 mM) for 30 min. The blot was exposed to lambda phosphatase (lanes 2 & 4) then probed with anti- SHP1 (C-terminal) antibody (lanes 1 & 2) or anti-SHP1 (Ser-591) antibody (lanes 3-6). The SHP1 (Ser-591) antibody was used in the presence of phospho-SHP1 (Ser-591) peptide (lane 5) or a non-specific phospho- serine peptide (lane 6).
Western blot analysis of ShcA expression in A431 cell lysate (lanes 1, 2, 3, & 4). The blots were probed with rabbit polyclonal anti-ShcA (SP1331) at 1:1000 (lane 1) or 1:4000 (lane 2) and mouse monoclonal ShcA (C-terminal region) at 1:1000 (lane 3) or 1:4000 (lane 4).Immunocytochemical labeling of ShcA in paraformaldehyde-fixed and NP-40-permeabilized A431 cells. The cells were labeled with mouse monoclonal (top) and rabbit polyclonal (bottom) ShcA antibodies, then the antibodies were detected using appropriate secondary antibodies conjugated to Cy3.

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