Anti-PP2A alpha + beta (methylation L309) antibody [2A10] (ab66597)

Overview

  • Product nameAnti-PP2A alpha + beta (methylation L309) antibody [2A10]
    See all PP2A alpha + beta primary antibodies
  • Description
    Mouse monoclonal [2A10] to PP2A alpha + beta (methylation L309)
  • Tested applicationsSuitable for: WB, ELISA, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Saccharomyces cerevisiae
  • Immunogen

    Synthetic peptide corresponding to Human PP2A alpha + beta conjugated to Keyhole Limpet Haemocyanin (KLH). A KLH-coupled peptide with the sequence: KLH-C-ßA-RRTPDYFL-OMe. The c-terminal sequence (as well as the carboxy-methylation) is absolutely conserved from human, mouse to yeast.

  • Positive control
    • This antibody gave a positive signal in Hek293 and PC12 whole cell lysates

Properties

Applications

Our Abpromise guarantee covers the use of ab66597 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
WB Use a concentration of 5 - 10 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
ELISA Use at an assay dependent dilution.
IP Use at an assay dependent dilution.

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 (methylation L309) antibody [2A10] images

  • All lanes : Anti-PP2A alpha + beta (methylation L309) antibody [2A10] (ab66597) at 10 µg/ml

    Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 36 kDa
    Observed band size : 36 kDa


    Exposure time : 20 minutes
  • Anti-PP2A alpha + beta (methylation L309) antibody [2A10] (ab66597) at 1/100 dilution + Lystae prepared from mouse embryonic fibroblast cells at 25 µg

    Secondary
    HRP-conjugatedsheep polyclonal to mouse IgG at 1/2000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 36 kDa
    Observed band size : 35 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 43 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute

    This image is a courtesy of Anonymous Abreview

    See Abreview

  • All lanes : Anti-PP2A alpha + beta (methylation L309) antibody [2A10] (ab66597) at 1/1000 dilution

    Lane 1 : No LCMT1 with 100% Licor Blocking Buffer for 30 minutes at 25°C.
    Lane 2 : LCMT1 at 0.02 µg/ml with 100% Licor Blocking Buffer for 30 minutes at 25°C.
    Lane 3 : LCMT1 at 0.1 µg/ml with 100% Licor Blocking Buffer for 30 minutes at 25°C.
    Lane 4 : LCMT1 at 0.5 µg/ml with 100% Licor Blocking Buffer for 30 minutes at 25°C.

    Secondary
    IRDye 800CW conjugated goat anti-mouse polyclonal at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 36 kDa

    This image is courtesy of an anonymous Abreview

    Detection method: Licor 10 minute exposure.
    Specific observed bands at 40kDa

    See Abreview

References for Anti-PP2A alpha + beta (methylation L309) antibody [2A10] (ab66597)

This product has been referenced in:
  • DeGrande ST  et al. Molecular mechanisms underlying cardiac protein phosphatase 2A regulation in heart. J Biol Chem 288:1032-46 (2013). WB . Read more (PubMed: 23204520) »
  • Benziane B  et al. Activation of AMP-activated Protein Kinase Stimulates Na+,K+-ATPase Activity in Skeletal Muscle Cells. J Biol Chem 287:23451-63 (2012). WB . Read more (PubMed: 22610379) »

See all 8 Publications for this product

Product Wall

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing
Sample Neurospora crassa Tissue lysate - whole (Hyphae)
Specification Hyphae
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Dr. Hila Shomin

Verified customer

Submitted Jul 28 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Sample Human Cell lysate - whole cell (HeLa)
Specification HeLa
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Oct 15 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Recombinant protein (PP2A A and C subunits)
Loading amount 0.5 µg
Specification PP2A A and C subunits
Gel Running Conditions Reduced Denaturing (10% Acrylamide gel)
Blocking step Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 10 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse Embryonic Fibroblast)
Loading amount 25 µg
Specification Mouse Embryonic Fibroblast
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Jul 27 2009

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