PP2A | Serine/threonine protein phosphatase 2A 59 kDa regulatory subunit B' gamma isoform

AS13 2747 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Oryza sativa

PP2A | Serine/threonine protein phosphatase 2A 59 kDa regulatory subunit B' gamma isoform

Product Information

Immunogen KLH-conjugated peptide, derived from Arabidopsis thaliana PP2A, UniProt: Q8RW96-1 , TAIR: AT4G15415

Host Rabbit

Clonality Polyclonal

Purity Immunogen affinity purified serum in PBS pH 7.4.

Format Lyophilized

Quantity 50 µg

Reconstitution For reconstitution add 50 µl of sterile water

Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.

Tested applications Immunolocalization (IL), Western blot (WB)

Recommended dilution 1 : 500 (IL), 1 : 1000 (WB)

Expected | apparent MW 59,1 kD

Reactivity

Confirmed reactivity Arabidopsis thaliana, Oryza sativa

Predicted reactivity Brassica olereacea, Pisum sativum
Species of your interest not listed? Contact us

Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application example

Total proteins were extracted from the indicated plant samples as previously described. 20 μg total proteins from: Arabidopsis thaliana cell suspension (1), 10 days old Arabidopsis thaliana seedlings (2), Oryza sativa cell suspension (3) (left panel) and Arabidopsis thaliana cell suspension (1), 10 days old Arabidopsis thaliana seedlings (2), Arabidopsis thaliana leafs (3) (right panel) were separated on 10 % SDS-PA gels, blotted onto PVDF membranes. Blots were blocked with 5 % milk powder in TBST for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody 0.25 μg/ml for 2h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:50 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was seconds.

Courtesy of Dr. Gábor Horváth, University of Szeged, Hungary

immunolocalization using anti-PP2A antibodies

immunolocalization using anti-PP2A antibodies

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

Additional information

Background

Background PP2A (Serine/threonine protein phosphatase 2A 59 kDa regulatory subunit B' gamma isoform) is required for the formation of the PP2A holoenzyme that negatively regulates brassinosteroid signaling by dephosphorylating and inactivating BRI1 in the cytoplasm and is involved in growth regulation and stress signaling.

Alternative names: AtB' gamma, PP2A, B' subunit, gamma isoform

Product citations

Confirmed reactivity:	Arabidopsis thaliana, Oryza sativa
predicted reactivity:	Brassica olereacea, Pisum sativum Species of your interest not listed? Contact us
not reactive in:	No confirmed exceptions from predicted reactivity are currently known
Picture (footer):	Application example Total proteins were extracted from the indicated plant samples as previously described. 20 μg total proteins from: Arabidopsis thaliana cell suspension (1), 10 days old Arabidopsis thaliana seedlings (2), Oryza sativa cell suspension (3) (left panel) and Arabidopsis thaliana cell suspension (1), 10 days old Arabidopsis thaliana seedlings (2), Arabidopsis thaliana leafs (3) (right panel) were separated on 10 % SDS-PA gels, blotted onto PVDF membranes. Blots were blocked with 5 % milk powder in TBST for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody 0.25 μg/ml for 2h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:50 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was seconds. 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
background:	PP2A (Serine/threonine protein phosphatase 2A 59 kDa regulatory subunit B' gamma isoform) is required for the formation of the PP2A holoenzyme that negatively regulates brassinosteroid signaling by dephosphorylating and inactivating BRI1 in the cytoplasm and is involved in growth regulation and stress signaling. Alternative names: AtB' gamma, PP2A, B' subunit, gamma isoform
calculated \| apparent molecular mass [kDa]:	59,1 kD
Clonality:	Polyclonal
Format:	Lyophilized
Host:	Rabbit
immunogen:	KLH-conjugated peptide, derived from Arabidopsis thaliana PP2A, UniProt: Q8RW96-1 , TAIR: AT4G15415
Purity:	Immunogen affinity purified serum in PBS pH 7.4.
Quantity:	50 µg
recommended dilution:	1 : 500 (IL), 1 : 1000 (WB)
Reconstitution:	For reconstitution add 50 µl of sterile water
storage:	Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications:	Immunolocalization (IL), Western blot (WB)

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