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.
Applications:Suitable for use in Western Blot, Immunocytochemistry, ELISA, Immunohistochemistry, Immunoprecipitation and Flow Cytometry.
Other applications have not been tested.
Recommended Dilutions:Western 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.
Immunocytochemistry: 5-15ug/ml shows positive staining of mitotic cells.
ELISA: 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).
Immunoprecipitation: Reported by an independent laboratory.
Optimal dilutions to be determined by the researcher.
Positive Control:Colcemid treated HeLa cell lysate or A431 mitotic cells fixed with 3.7% paraformaldehyde.
Storage and Stability:May be stored at 4°C for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20°C.
Aliquots are stable for 12 months after receipt.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：200ug
Host：mouse
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by Protein G affinity chromatography.
Form：Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, 30% glycerol.
Specificity：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.
Clone # USB：0.T.181
Isotype：IgG1
Calc Applications Abbrev：E FC IC IHC IP WB
Calc Crossreactivity：Hu
Immunogen：Mitotic human HeLa cell cytosolic lysate.
