Product nameAnti-Mitotic proteins antibody [MPM-2]
DescriptionMouse monoclonal [MPM-2] 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. The clone number has been updated from (0.T.181) to (MPM-2) both clone numbers name the same antibody clone.
Tested applicationsSuitable for: IHC-P, ICC/IF, IP, WB, ICC, ELISAmore details
Species reactivityReacts with: Human
Predicted to work with: a wide range of other species
Tissue/ cell preparation (Mitotic human HeLa cell cytosolic lysate).
- Colcemid treated HeLa cell lysate.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.05% Sodium azide
Constituents: 0.184% Tris glycine, 0.87% Sodium chloride
Concentration information loading...
PurityProtein G purified
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.
|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).|
Cellular localizationThe phosphoproteins have been found in the cytoplasm of mitotic cells and near centrosomes, kinetochores, and midbodies of the microtubule organizing centers.
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.
This product has been referenced in:
- Ruppert JG et al. HP1a targets the chromosomal passenger complex for activation at heterochromatin before mitotic entry. EMBO J 37:N/A (2018). Flow Cyt . Read more (PubMed: 29467217) »
- 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) »