PP2A | Serine/threonine protein phosphatase 2A 59 kDa regulatory subunit B' gamma isoform
AS13 2747 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Oryza sativa
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Courtesy of Dr. Gábor Horváth, University of Szeged, Hungary
Immunolocalization of PP2A on Arabidopsis thaliana, MM1 (upper panel) and Oryza sativa ssp. japonica cv ‘Unggi 9’ (lower panel) plant suspension cultures. DyLight488 (Agrisera AS10 831) anti-rabbit antibody is used as fluorescent conjugated secondary antibody (green). DAPI is used as nuclear marker (red). Merged transmission images (differential interference contrast) are shown at the last column. Scalebars are 10 µm.
Labeling and detection protocol
Fixation (30mins) 4% paraformaldehyde in PBS (pH 7.4) with 0.05% TritonX-100. 3x5min PBS wash. Cell wall digestion (30mins): 1% Cellulase, 0.5% Pectinase in 0.5% (w/v) MES buffer (pH 5.6). 2x5min PBS wash.
Immobilization of cells (10mins): Cells in PBS were settled on poly-L-Lysine coated coverslips, excess PBS removed without air drying the cells. Membrane permeabilization (10mins): 0.5% TritonX-100 in PBS. 3x5min PBS wash. Blocking (10mins): 5% Fish gelatin in PBS. Primary antibody incubation (16 h at 4°C or 1h at 37°C): Agrisera (AS13 2747) rabbit anti PP2A antibody diluted 1:500 in blocking buffer. 4x5min blocking buffer wash. Secondary antibody incubation (1h at 37°C): Agrisera (AS10 831) chicken anti-rabbit DyLight 488 antibody diluted 1:200 in blocking buffer. 3x5min PBS wash. Nuclear counterstaining (5mins): 200ng/µl DAPI in PBS. Brief PBS wash. Mounting: Fluoromount G mounting medium was used to mount coverslips onto glass slides. Imaging: Olympus FV1000 confocal microscope with 40x (NA1.3) oil immersion objective.
Courtesy of Dr. Ferhan Ayaydin, Cellular Imaging Laboratory, Biological Research Center, Szeged, Hungary
Alternative names: AtB' gamma, PP2A, B' subunit, gamma isoform