MPM2, Ser/Thr, Pro, phosphorylated (Mitotic Protein Monoclonal#2)

Upon entry into M (Mitosis) phase, many proteins are phosphorylated either directly or indirectly by M-phase-promoting factor (MPF). The MPM2 monoclonal antibody binds to a phospho aa-containing epitope (peptides containing LTPLK and FTPLQ domains) present on more than 50 proteins of M-phase eukaryotic cells. This antibody recognizes a variety of proteins that are phosphorylated during mitosis.\n\nApplications:\nSuitable for use in Western Blot, Immunocytochemistry, ELISA, Immunohistochemistry, Immunoprecipitation and Flow Cytometry. Other applications have not been tested. \n\nRecommended Dilutions:\nWestern Blot: 0.5-2ug/ml detects mitosis-associated proteins in colcemid treated (0.05ug/ml) human HeLa carcinoma cells. Colcemid-treated HeLa cell lysates were resolved by electrophoresis, transferred to nitrocellulose and probed with M4685. Proteins were visualized using an IgG (HRP) goat anti-mouse antibody and ECL.\nImmunocytochemistry: 5-15ug/ml shows positive staining of mitotic cells.\nELISA: Used in a non-radioactive, dissociation-enhanced time resolved fluoroimmunoassay (DELPHIA). 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: Serine/Threonine Kinase Substrate Peptide, Recombinant, Human (Biotin).\nImmunoprecipitation: Reported by an independent laboratory.\nOptimal dilutions to be determined by the researcher.\n\nPositive Control: \nColcemid treated HeLa cell lysate or A431 mitotic cells fixed with 3.7% paraformaldehyde.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

M4685

Size

200ug

Applications

ELISA, FC, ICC, IHC, IP, WB

Hosts

Mouse

Reactivities

Hum

Form

Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, 30% glycerol.

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG1

References

1. Hresko, R.C., et al., J. Biol. Chem. 278: 21,615-21,622 (2003). 2. Escargueil, A.E., et al., J. Biol. Chem. 275: 34,710-34,718 (2000). 3. Andreassen, P.R., et al., Mol. Biol. Cell 12: 1315-1328 (2001). 4. Michel, L.S., et al., Nature 409: 355-359 (2001). 5. Vincent, I., et al., J. Neurosci. 17: 3588-3598 (1997). 6. Yaffe, M.B., et al., Science 278: 1957-1960 (1997). 7. Harper, J.D., et al., Eur. J. Cell Biol. 51: 272-278 (1990). 8. Kuang, J., et al., Proc. Natl. Acad. Sci. USA 86: 4982-4986 (1989). 9. Kuang, J., et al., Mol. Biol. Cell 5: 135-145 (1994). 10. Westendorf, J.M., et al., Proc. Natl. Acad. Sci. USA 91: 714-718 (1994). 11. Albert, A.L., et al., BMC Cell Biol. 5: 22 (2004). 12. Rosse, C., et al., J. Biol. Chem. 278: 30,597-30,604 (2003).

Additional Info

Recognizes human MPM2. Reacts with phosphorylated serines or threonines when followed by a proline. Recognizes 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 MPM2 varies from species to species and with the cell type. Species Crossreactivity: animals and plants.