Name: Anti-PP2A alpha + beta antibody [YE351]
Brand: RabMAb
SKU: ab32065
Rating: 5 (1 reviews)

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Western blot - 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

Carrier Free

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.
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.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

YE351
Isotype

IgG
Research areas

Alternative Versions
- Anti-PP2A alpha + beta antibody [YE351] - BSA and Azide free (ab232450)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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
IP		1/80.
WB	(1)	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.

Notes
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.

Application notes

Is unsuitable for Flow Cyt or ICC/IF.

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
modifications

Reversibly 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.
Target information above from: UniProt accession P67775 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
- Omim: 176915 Human
- Omim: 176916 Human
- SwissProt: P67774 Cow
- SwissProt: Q0P594 Cow
- SwissProt: P62714 Human
- SwissProt: P67775 Human
- SwissProt: P62715 Mouse
- SwissProt: P63330 Mouse
- SwissProt: P62716 Rat
- SwissProt: P63331 Rat
- Unigene: 105818 Human
- Unigene: 260288 Mouse
- Unigene: 1271 Rat
see all
Alternative names
- PP2A A antibody
- PP2A alpha antibody
- PP2A B antibody
- PP2A beta antibody
- PP2A-alpha antibody
- PP2A-beta antibody
- PP2AA_HUMAN antibody
- PP2Aalpha antibody
- PP2AB_HUMAN antibody
- PP2Abeta antibody
- PP2Ac antibody
- PP2CA antibody
- PP2Calpha antibody
- PP2CB antibody
- PPP2CA antibody
- PPP2CB antibody
- Protein phosphatase 2 catalytic subunit alpha isoform antibody
- Protein phosphatase 2 catalytic subunit beta isoform antibody
- Protein phosphatase type 2A catalytic subunit antibody
- Replication protein C antibody
- RP C antibody
- RP-C antibody
- RPC antibody
- Serine/threonine protein phosphatase 2A catalytic subunit alpha isoform antibody
- Serine/threonine protein phosphatase 2A catalytic subunit beta isoform antibody
- Serine/threonine-protein phosphatase 2A catalytic subunit alpha isoform antibody
see all

Western blot - Anti-PP2A alpha + beta antibody [YE351] (ab32065)

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 seconds

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-PP2A alpha + beta antibody [YE351] (ab32065)

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 seconds

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-PP2A alpha + beta antibody [YE351] (ab32065)

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 seconds

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-PP2A alpha + beta antibody [YE351] (ab32065)

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 seconds

Blocking and dilution buffer: 5% NFDM /TBST.
Western blot - Anti-PP2A alpha + beta antibody [YE351] (ab32065)

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 second

Blocking and dilution buffer: 5% NFDM /TBST.
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.
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
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.
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.
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.
Immunoprecipitation - Anti-PP2A alpha + beta antibody [YE351] (ab32065)

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.
Dot Blot - Anti-PP2A alpha + beta antibody [YE351] (ab32065)

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.
Anti-PP2A alpha + beta antibody [YE351] (ab32065)

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.

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab32065? Please let us know so that we can cite the reference in this datasheet.

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

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Application

Western blot

Sample

Mouse Tissue lysate - whole (mouse brain)

Gel Running Conditions

Reduced Denaturing (4-20% gel)

Loading amount

20 µg

Specification

mouse brain

Blocking step

(agent) for 1 hour(s) and 0 minute(s) · Concentration: 100%

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Apr 12 2021

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Epitope

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (5)

Customer reviews and Q&As

Western blot abreview for Anti-PP2A alpha + beta antibody [YE351]

Post-translational
modifications