Cell Signaling Technology

Product Pathways - Ca / cAMP / Lipid Signaling

Phospho-PKD/PKCμ (Ser744/748) Antibody #2054

Applications Reactivity Sensitivity MW (kDa) Source
W H M R Mk Endogenous 115 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-PKD/PKCμ (Ser744/748) Antibody detects PKD1/PKCμ only when dually phosphorylated at serines 744 and 748. This antibody may also cross-react with isoforms PKD2 and PKD3/PKCη in some species.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser744/748 of mouse PKD. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated, PDGF-treated (50 ng/ml) or TPA-treated (0.2 µM), using Phospho-PKD/PKCµ (Ser744/748) Antibody (upper) or PKD/PKCµ Antibody #2052 (lower).

Background

Activation of PKC is one of the earliest events in a cascade leading to a variety of cellular responses such as secretion, gene expression, proliferation and muscle contraction (1,2). Protein kinase D (PKD), also called PKCμ, is a serine/threonine kinase whose activation is dependent on the phosphorylation of two activation loop sites, Ser744 and Ser748, via a PKC-dependent signaling pathway (3-5). In addition to the two activation loop sites, the carboxy-terminal Ser916 has been identified as an autophosphorylation site for PKD/PKCμ. Phosphorylation at Ser916 correlates with PKD/PKCμ catalytic activity (6).

  1. Nishizuka, Y. (1984) Nature 308, 693-698.
  2. Keranen, L.M. (1995) Curr. Biol. 5, 1394-1403.
  3. Valverde, A.M. et al. (1994) Proc. Natl. Acad. Sci. 91, 8572-8576.
  4. Johannes, F.J. et al. (1994) J. Biol. Chem. 269, 6140-6148.
  5. Iglesias, T. et al. (1998) J. Biol. Chem. 273, 27662-27667.
  6. Matthews, S.A. et al. (1999) J. Biol. Chem. 274, 26543-26549.
  7. Wood, C.D. et al. (2005) J Biol Chem 280, 6245-51.
  8. Amadesi, S. et al. (2009) J Comp Neurol 516, 141-56.

Application References

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