Recombinant Anti-PP2A alpha + beta antibody [YE351] (ab32065)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [YE351] to PP2A alpha + beta
- Suitable for: IP, WB, IHC-P, Dot blot
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-PP2A alpha + beta antibody [YE351]
See all PP2A alpha + beta primary antibodies -
Description
Rabbit monoclonal [YE351] to PP2A alpha + beta -
Host species
Rabbit -
Specificity
This antibody can bind both alpha and beta form of PP2A. Mouse samples have been successfully tested in WB, but IHC-P sections from various mouse tissues showed negative results. -
Tested applications
Suitable for: IP, WB, IHC-P, Dot blotmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Cow, Pig -
Immunogen
Synthetic peptide within Human PP2A alpha + beta (N terminal). The exact sequence is proprietary.
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Epitope
ab32065 reacts with an epitope located in the N terminal region of PP2A alpha. -
Positive control
- WB: A431 cell lysate and NIH 3T3 cell lysate. Dot Blot: PP2A-alpha peptide and PP2A-beta peptide.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
YE351 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32065 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IP |
1/80.
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WB | (1) |
1/500. Predicted molecular weight: 36 kDa.
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Dot blot |
Use at an assay dependent concentration.
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Notes |
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IP
1/80. |
WB
1/500. Predicted molecular weight: 36 kDa. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Dot blot
Use at an assay dependent concentration. |
Target
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Function
PP2A can modulate the activity of phosphorylase B kinase casein kinase 2, mitogen-stimulated S6 kinase, and MAP-2 kinase. Cooperates with SGOL2 to protect centromeric cohesin from separase-mediated cleavage in oocytes specifically during meiosis I (By similarity). Can dephosphorylate SV40 large T antigen and p53/TP53. Dephosphorylates SV40 large T antigen, preferentially on serine residues 120, 123, 677, and perhaps 679. The C subunit was most active, followed by the AC form, which was more active than the ABC form, and activity of all three forms was strongly stimulated by manganese, and to a lesser extent by magnesium. Dephosphorylation by the AC form, but not C or ABC form is inhibited by small T antigen. Activates RAF1 by dephosphorylating it at 'Ser-259'. -
Sequence similarities
Belongs to the PPP phosphatase family. PP-1 subfamily. -
Post-translational
modificationsReversibly methyl esterified on Leu-309. Carboxyl methylation may play a role in holoenzyme assembly, enhancing the affinity of the PP2A core enzyme for some, but not all, regulatory subunits. It varies during the cell cycle.
Phosphorylation of either threonine (by autophosphorylation-activated protein kinase) or tyrosine results in inactivation of the phosphatase. Auto-dephosphorylation has been suggested as a mechanism for reactivation. -
Cellular localization
Cytoplasm. Nucleus. Chromosome > centromere. Cytoplasm > cytoskeleton > spindle pole. In prometaphase cells, but not in anaphase cells, localizes at centromeres. During mitosis, also found at spindle poles. Centromeric localization requires the presence of SGOL2. - Information by UniProt
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Database links
- Entrez Gene: 5515 Human
- Entrez Gene: 5516 Human
- Entrez Gene: 19052 Mouse
- Entrez Gene: 19053 Mouse
- Entrez Gene: 397136 Pig
- Entrez Gene: 397656 Pig
- Entrez Gene: 24672 Rat
- Entrez Gene: 24673 Rat
see all -
Alternative names
- PP2A A antibody
- PP2A alpha antibody
- PP2A B antibody
see all
Images
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All lanes : Anti-PP2A alpha + beta antibody [YE351] (ab32065) at 1/50000 dilution
Lane 1 : A431 whole cell lysate
Lane 2 : A549 whole cell lysate
Lane 3 : Caco-2 whole cell lysate
Lane 4 : T47D whole cell lysate
Lane 5 : C2C12 whole cell lysate
Lane 6 : HeLa whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Exposure time: 15 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-PP2A alpha + beta antibody [YE351] (ab32065) at 1/50000 dilution
Lane 1 : Jurkat whole cell lysate
Lane 2 : U-937 whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Exposure time: 5 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-PP2A alpha + beta antibody [YE351] (ab32065) at 1/100000 dilution
Lane 1 : Human fetal heart tissue lysate
Lane 2 : Human fetal kidney tissue lysate
Lane 3 : Human fetal spleen tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Exposure time: 30 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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Anti-PP2A alpha + beta antibody [YE351] (ab32065) at 1/100000 dilution + Human fetal brain tissue lysate at 10 µg
Secondary
HRP-conjugate anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Exposure time: 15 secondsBlocking and dilution buffer: 5% NFDM /TBST.
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All lanes : Anti-PP2A alpha + beta antibody [YE351] (ab32065) at 1/100000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse heart tissue lysate
Lane 3 : Mouse kidney tissue lysate
Lane 4 : Mouse spleen tissue lysate
Lane 5 : Rat brain tissue lysate
Lane 6 : Rat heart tissue lysate
Lane 7 : Rat kidney tissue lysate
Lane 8 : Rat spleen tissue lysate
Lane 9 : C6 whole cell lysate
Lane 10 : Raw264.7 whole cell lysate
Lane 11 : PC-12 whole cell lysate
Lane 12 : NIH/3T3 whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Exposure time: 1 secondBlocking and dilution buffer: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PP2A alpha + beta antibody [YE351] (ab32065)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labeling PP2A alpha + beta with ab32065 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Cytoplasm and weak nucleus staining is visible in lymphocytes of human tonsil.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PP2A alpha + beta antibody [YE351] (ab32065)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labelling PP2A alpha + beta with ab32065 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Cytoplasm staining is visible in epithelial cells of mouse kidney
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PP2A alpha + beta antibody [YE351] (ab32065)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling PP2A alpha + beta with ab32065 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Cytoplasm and weak nucleus staining is visible in epithelial cells of rat kidney.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PP2A alpha + beta antibody [YE351] (ab32065)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse heart tissue labelling PP2A alpha + beta with ab32065 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Cytoplasm staining is visible in mouse heart.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PP2A alpha + beta antibody [YE351] (ab32065)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat heart tissue labelling PP2A alpha + beta with ab32065 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Cytoplasm and nucleus staining is visible on rat heart.
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ab32065 at 1/80 immunoprecipitating PP2A alpha + beta in HeLa whole cell lysate.
Lane 1 (input): HeLa whole cell lysate (10µg)
Lane 2 (+): ab32065 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32065 in HeLa (human cervix adenocarcinoma) whole cell lysate.
For western blotting, ab32065 (1/500) ab131366 VeriBlot for IP Detection Reagent (HRP) was used for detection (1/10000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Observed band: 37kDa.
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Dot blot analysis of PP2A-alpha peptide (Lane 1) and PP2A-beta peptide (Lane 2) labelling PP2A alpha+beta with ab32065 at a dilution of 1/1000. A HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody at a dilution of 1/1000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (5)
ab32065 has been referenced in 5 publications.
- Yuan P et al. Proteomics reveals the potential mechanism of Tanshinone IIA in promoting the Ex Vivo expansion of human bone marrow mesenchymal stem cells. Regen Ther 21:560-573 (2022). PubMed: 36475023
- Albadrani M et al. Exogenous PP2A inhibitor exacerbates the progression of nonalcoholic fatty liver disease via NOX2-dependent activation of miR21. Am J Physiol Gastrointest Liver Physiol 317:G408-G428 (2019). PubMed: 31393787
- Tang S et al. Hydrogen peroxide redistributes the localization of protein phosphatase methylesterase 1. Life Sci 213:166-173 (2018). PubMed: 30340029
- Park HJ et al. Protein Phosphatase 2A and Its Methylation Modulating Enzymes LCMT-1 and PME-1 Are Dysregulated in Tauopathies of Progressive Supranuclear Palsy and Alzheimer Disease. J Neuropathol Exp Neurol 77:139-148 (2018). PubMed: 29281045
- Langlois B et al. LRP-1 promotes cancer cell invasion by supporting ERK and inhibiting JNK signaling pathways. PLoS One 5:e11584 (2010). WB . PubMed: 20644732