Overview

  • Product name

    Mouse PP2A-alpha ELISA Kit
    See all PP2A-alpha kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Cell extract 8 5.5%
    Inter-assay
    Sample n Mean SD CV%
    Cell extract 5 7.3%
  • Sample type

    Cell culture extracts, Tissue Extracts, Tissue Homogenate
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    66.2 pg/ml
  • Range

    85.9 pg/ml - 5500 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 102 98% - 106%
    Tissue Extracts 97 92% - 100%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Product overview

    Abcam’s PP2A-alpha in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of PP2A-alpha protein in human cell and tissue extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    PP2A-alpha is a highly conserved heterotrimeric phosphatase for microtubule-associated proteins. Additionally, PP2A-alpha targets include proteins in oncogenic signalling cascades, including AKT, MEK and Raf.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse PP2A-alpha Capture Antibody 1 x 600µl
    10X Mouse PP2A-alpha Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI 1 x 6ml
    Mouse PP2A-alpha Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • 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.
  • 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. 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
  • Alternative names

    • PP2A alpha
    • PP2A-alpha
    • PP2AA_HUMAN
    • PP2Aalpha
    • PP2Ac
    • PP2CA
    • PP2Calpha
    • PPP2CA
    • Protein phosphatase 2 (formerly 2A) catalytic subunit alpha isoform
    • Protein phosphatase 2 catalytic subunit alpha isoform
    • Protein phosphatase 2, catalytic subunit, alpha isozyme
    • Protein phosphatase 2A catalytic subunit, alpha isoform
    • Replication protein C
    • RP C
    • RP-C
    • RPC
    • Serine/threonine protein phosphatase 2A catalytic subunit alpha isoform
    • Serine/threonine-protein phosphatase 2A catalytic subunit alpha isoform
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab209878 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of PP2A-alpha were measured in duplicate and interpolated from the PP2A-alpha standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PP2A-alpha concentration was determined to be 4,273 pg/mL in NIH-3T3 extract and 2,719 pg/mL in mouse skeletal muscle extract.

  • The concentrations of PP2A-alpha in PC12 and HeLa extract were measured in duplicate, interpolated from the mouse PP2A-alpha standard curves and corrected for sample dilution. Undiluted samples are PC12 and HeLa extract based on a 10 μg/mL extract load. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).  The mean PP2A-alpha concentration was determined to be 4,956 pg/mL in PC12 extract and 4,395 pg/mL in HeLa extract.

Protocols

References

ab209878 has not yet been referenced specifically in any publications.

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