Myelin Basic Protein, Dephosphorylated, Bovine (MBP)

In vitro, myelin basic protein (MBP) is a substrate for phosphorylation by several different protein kinases including MAPK, PKA, calmodulin-dependent protein kinase, PKC and phosphorylase kinase. Even highly specific protein kinases such as Raf1, MEK and MEKK can utilize MBP as an alternative substrate. Myelin basic protein (MBP) is a single-chain, flexible polypeptide of about 18.5kD existing in its natural environment as an extrinsic and loosely bound component of the cytoplasmic portion of the myelin membrane sandwich. Ultrastructural immunocytochemistry with anti-myelin basic protein has shown that MBP is localized in the compact myelin sheath. MBP has not been demonstrated in rough endoplasmic reticulum, lysosomes or any other cytoplasmic organelles. MBP serves as a marker for oligodendrocytes and Schwann cells. MBP is also a marker for malignant schwannomas and appears to play a significant role in the etiology of multiple sclerosis and experimental autoimmune encephalomyelitis.\n\nProtein Kinase Assay: \nDephosphorylated MBP incorporated 32P-phosphate in radioactive kinase reactions using MAPK2/Erk2. Phosphorylated in a non-radioactive kinase assay using M2362-06 MAP Kinase 2, Recombinant, Murine. \nMBP phosphorylation was detected by immunoblot analysis usingM9758-05 Myelin Basic Protein, phosphorylated (MBP). No detectable endogenous phosphorylation on MBP was seen as determined by immunoblotting 1ug of the dephosphorylated MBP with 1ug/ml M9758-05 Myelin Basic Protein, phosphorylated (MBP). \n\nPhosphatase Activity: \nWhen incubated with pNPP for 15 minutes, 250ug had no detectable levels of active phosphatase. \n\nImmunoblot Analysis: 800ng MBP incubated without or with MAPK2 and probed with M9758-06 MBP Dephosphorylated, Bovine (MBP) (1ug/ml). Recognizes MBP at ~21kD when incubated with MAPK2.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. 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.

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SPECIFICATIONS

Catalog Number

M9758-06

Size

5mg

Applications

WB

Form

Supplied as a liquid in 10mM MOPS, pH 7.0, 128uM MnCl2, 641uM EDTA, 1.134ug inactive lambda phosphatase.

Purity

Purified from bovine brain using HPLC and de-phosphorylated using Lambda protein phosphatase. Lambda protein phosphatase was inactivated, in the presence of 10mM sodium EDTA, at 65 degrees C for one hour. Purity was assessed at 95% using SDS-PAGE and Coomassie blue staining of 1ug of product. Developed for use in radioactive and non-radioactive kinase assays.

References

1. Cicirelli, et al., J. Biol. Chem. 263: 2009, 1988. 2. Martenson, et al., J. Biol. Chem. 258: 930, 1983. 3. Ulmer, et al., J. Biol. Chem. 262: 1748, 1987.\nGeneral references:\n1 Day, E. D. and Potter, N. T (1986) J. Neuroimmunol. 10, 289. 2 Hashim, G. A. (1978) Immunol. Rev. 39, 60. 3 Golo, M. (1987) Histochemistry 87, 201. 4 Mogollon, R, et al. (1984) Cancer 53, 1190. 5 Whitaker, J. N. (1984) Ann. N.Y. Acad. Sci. 436, 140. 6 Eidinoff, H. (1988) Med. Hypotheses 26, 103. 7 tar Meulen, V. (1988) Ann. N.Y. Acad. Sci. 540, 202. 8 Kerlero de Rosbo, N. et al. (1985) J. Neuroimmunol. 9, 349. 9 Sires, L. R. et al. (1981) Science 214, 87.