Recombinant
RabMAb

Recombinant Anti-PP2A alpha + beta antibody [E155] - BSA and Azide free (ab226007)

Overview

  • Product name
    Anti-PP2A alpha + beta antibody [E155] - BSA and Azide free
    See all PP2A alpha + beta primary antibodies
  • Description
    Rabbit monoclonal [E155] to PP2A alpha + beta - BSA and Azide free
  • Host species
    Rabbit
  • Specificity
    The immunogen sequence for ab32104 has 100% homology with the PP2A-beta protein.
  • Tested applications
    Suitable for: Flow Cyt, IHC-P, WB, ICC/IF, IP, Dot blotmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Zebrafish
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human PP2A alpha + beta aa 250-350.

  • Positive control
    • IHC-P: Human pancreas tissue. WB: lysate of A431 cells (serum starved overnight) treated with EGF (100ng/ml) for 5-10 minutes at 37C (BD Biosciences).
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®). 

    ab226007 is a PBS only buffer version of ab32104, containing no BSA or sodium azide, ideal for antibody labeling. Please refer to ab32104 for information on protocols, dilutions, and image data.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab226007 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
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

IHC-P Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 35 kDa.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Dot blot Use at an assay dependent concentration.

Target

  • 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.
  • Information by UniProt
  • Database links
  • Alternative names
    • PP2A A antibody
    • PP2A alpha antibody
    • PP2A B antibody
    • PP2A beta antibody
    • PP2A-alpha antibody
    • PP2A-beta antibody
    • PP2AA_HUMAN antibody
    • PP2AB_HUMAN antibody
    • PP2Abeta 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
    • Protein phosphatase type 2A catalytic subunit 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

Images

  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling PP2A alpha + beta with purified ab32104 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32104).

  • Dot blot was performed using ab32104 at 1/1000 dilution.Goat Anti-Rabbit IgG,(H+L),HRP conjugated (ab97051) was used as secondary antibody at 1/100000 dilution

    Lane 1:PP2A alpha + beta  phospho peptide

    Lane 2:PP2A alpha + beta non-phospho peptide

    Blocking and Diluting buffer and concentration:5% NFDM/TBST

    Exposure time:10 seconds

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32104).

  • ICC/IF image of ab32104 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32104, 5µg/ml) overnight at +4°C. The secondary antibody (green)�was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32104).

  • This IHC data was generated using the same anti-PP2A alpha + beta antibody clone [E155] in a different buffer formulation (cat# ab32104).

    ab32104 (2µg/ml) staining PP2A alpha  in human pancreas using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of the islet of Langerhans.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

References

This product has been referenced in:
  • Inoue D  et al. SETBP1 mutations drive leukemic transformation in ASXL1-mutated MDS. Leukemia 29:847-57 (2015). Read more (PubMed: 25306901) »
  • Cristóbal I  et al. PP2A inhibition is a common event in colorectal cancer and its restoration using FTY720 shows promising therapeutic potential. Mol Cancer Ther 13:938-47 (2014). Human . Read more (PubMed: 24448818) »
See all 18 Publications for this product

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