Anti-Mitotic proteins antibody [0.T.181] (ab14581)

Overview

  • Product nameAnti-Mitotic proteins antibody [0.T.181]
  • Description
    Mouse monoclonal [0.T.181] to Mitotic proteins
  • SpecificityRecognizes a phosphorylated epitope (S/T)P found in phosphoproteins such as MAP2, HSP70, cdc25 and DNA topoisomerase IIa, most of which are phosphorylated at the onset of mitosis. The number of phosphoproteins recognized by MPM-2 varies from species to species and with the cell type.
  • Tested applicationsSuitable for: IHC-P, ICC/IF, IP, WB, ICC, ELISAmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: a wide range of other species
  • Immunogen

    Tissue/ cell preparation (Mitotic human HeLa cell cytosolic lysate).

  • Positive control
    • Colcemid treated HeLa cell lysate.
  • General notesFor maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Properties

Applications

Our Abpromise guarantee covers the use of ab14581 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
IHC-P Use a concentration of 2 µg/ml.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 92 kDa.
ICC Use a concentration of 5 - 15 µg/ml.
ELISA Use at an assay dependent concentration. This antibody was used in a non-radioactive, dissociation-enhanced time resolved fluoroimmunoassa.Active enzymes were used to phosphorylate substrate peptides. The phosphorylated peptides were detected using MPM2 in conjunction with a europium (Eu3+) labeled secondary antibody. The phosphorylated peptides detected include: S1000-50 Serine Kinase Substrate Peptide (Biotin) (RRRAPLSP-N-Biotin) and S1000-80 Serine/Threonine Kinase Substrate Peptide, Recombinant, Human (Biotin).

Target

  • Cellular localizationThe phosphoproteins have been found in the cytoplasm of mitotic cells and near centrosomes, kinetochores, and midbodies of the microtubule organizing centers.

Anti-Mitotic proteins antibody [0.T.181] images

  • Ab14581 staining human normal skin. Staining is localised to the nucleus.
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • ab14581 staining Mitotic proteins - Mitosis Marker in Human Saos-2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton in PBS and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/500) for 1 hour. An Alexa Fluor® 594-conjugated Goat anti-mouse IgG polyclonal (1/250) was used as the secondary antibody.

    See Abreview

References for Anti-Mitotic proteins antibody [0.T.181] (ab14581)

This product has been referenced in:
  • Machicoane M  et al. SLK-dependent activation of ERMs controls LGN-NuMA localization and spindle orientation. J Cell Biol 205:791-9 (2014). Read more (PubMed: 24958772) »
  • Arai A  et al. RECQL1 and WRN Proteins Are Potential Therapeutic Targets in Head and Neck Squamous Cell Carcinoma. Cancer Res 71:4598-607 (2011). Read more (PubMed: 21571861) »

See all 7 Publications for this product

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (saos-2 human osteosarcoma)
Specification saos-2 human osteosarcoma
Fixative Paraformaldehyde
Permeabilization Yes - pbs triton 0.25%
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: rt°C
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Submitted Apr 19 2012

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