MARCKS (phospho Ser152 / 156) Antibody

Myristoylated Alanine-Rich C Kinase Substrate (MARCKS) is a major substrate for phosphorylation by protein kinase C (PKC) . The phosphorylation of Ser152/156 can be used as a measure of PKC activation. Moreover, MARCKS is a member of a family of calmodulin binding proteins. Phosphorylation of Ser152/156 modulates the binding of MARCKS to calmodulin.

MARCKS (Ser152/156) polyclonal antibody was raised against a synthetic phosphopeptide corresponding to amino acid residues surrounding the phospho-Ser152 and phospho-Ser156 of rat MARCKS.

For long term storage –80˚ C is recommended, but shorter term storage at –20˚ C is also acceptable as aliquots may be taken without freeze/thawing due to the presence of 50% glycerol. Stable for one year.

Immunolabeling of the 87kDa band in rat brain homogenate is blocked by the Ser152/156 phosphopeptide but not by the non-phosphopeptide. The antibody is purified by sequential chromatography on phospho- and non-phosphopeptide affinity columns. Antibody dilutions and tissue load should be based on tissue type and expected phosphorylation state. Initial recommended range of dilutions: 1:500 to 1:2000. Applications include Dot Blots (DB) and Western Blot (WB) . Suitability for Immunohistochemistry (IHC) not yet determined. Human, mouse, rat, and Xenopus have 100% amino acid sequence identity with the antigen used to raise the antibody. When internally tested under ideal conditions the working dilutions were 1:1000 for DB and WB.

Palmer, R.H., Schö nwasser, D.C., Rahman, D., Pappin, D.J.C., Herget, T., and Parker, P.J., "PRK1 phosphorylates MARCKS at the PKC sites: Serine 152, serine 156 and serine 163, " FEBS Letters, 378 (1996) 281 - 285.

Spizz, G. and Blackshear, P.J., "Protein kinase C-mediated phosphorylation of the myristoylated alanine-rich C-kinase substrate protects it from specific proteolytic cleavage, " J. Biol. Chem., 271 (1996) 553 - 562.

Thelen, M., Rosen, A., Nairn, A.C., and Aderem, A., "Regulation by phosphorylation of reversible association of a myristoylated protein kinase C substrate with the plasma membrane, " Nature (London), 351 (1991) 320 - 322.

Hartwig, J.H., Thelen, M., Rosen, A., Janmey, P.A., Nairn, A.C., and Aderem, A., "MARCKS is an actin filament crosslinking protein regulated by protein kinase C and calcium-calmodulin, " Nature (London), 356 (1992) 618 - 622.