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

Overview

  • Product nameAnti-PP2A alpha + beta antibody [YE351]
    See all PP2A alpha + beta primary antibodies
  • Description
    Rabbit Monoclonal [YE351] to PP2A alpha + beta
  • SpecificityThis 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 applicationsSuitable for: IP, WB, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Pig
  • Immunogen

    corresponding to Human PP2A alpha + beta (N terminal).

  • Epitopeab32065 reacts with an epitope located in the N terminal region of PP2A alpha.
  • Positive control
    • A431 cell lysate and NIH 3T3 cell lysate.
  • General notes

    This product is a recombinant rabbit monoclonal antibody. 

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

Properties

Applications

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 1/500. Predicted molecular weight: 36 kDa.
IHC-P 1/50 - 1/100.
  • Application notesIs unsuitable for Flow Cyt or ICC/IF.
  • Target

    • FunctionPP2A 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 similaritiesBelongs 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 localizationCytoplasm. 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
    • Database links
    • Alternative names
      • PP2A A antibody
      • PP2A alpha antibody
      • PP2A B antibody
      • PP2A beta antibody
      • PP2A-alpha antibody
      • PP2AA_HUMAN antibody
      • PP2Ac antibody
      • PP2CB antibody
      • PPP2CA antibody
      • PPP2CB antibody
      • Protein phosphatase 2 catalytic subunit alpha isoform antibody
      • Protein phosphatase 2 catalytic subunit beta isoform antibody
      • Replication protein C antibody
      • RP C antibody
      • RP-C 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

    Anti-PP2A alpha + beta antibody [YE351] images

    • 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
      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.

    • 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
      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.

    • 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
      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.

    • 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.

    • 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
      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) analysis of human tonsil 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 lymphocytes of human tonsil.

    • 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) 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) 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) 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.

    • 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 (HRP) was used as the secondary antibody (1/10000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

      Observed band: 37kDa.

       

    • 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.

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

    This product has been referenced in:
    • Langlois B  et al. LRP-1 promotes cancer cell invasion by supporting ERK and inhibiting JNK signaling pathways. PLoS One 5:e11584 (2010). WB . Read more (PubMed: 20644732) »

    See 1 Publication for this product

    Product Wall

    Thank you for your patience in awaiting our reply while we gathered our data on this. It does not appear that this has been specifically tested for our products but comparing the immunogens of our PP2CA antibodies there is a strong possibility that eac...

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"