Cell Signaling Technology

Product Pathways - Neuroscience

Phospho-Tyrosine Hydroxylase (Ser40) Antibody #2791

Applications Reactivity Sensitivity MW (kDa) Source
W IF-IC R (H) (M) Endogenous 55-60 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat
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-Tyrosine Hydroxylase (Ser40) Antibody detects endogenous levels of tyrosine hydroxylase only when phosphorylated at serine 40.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to the sequence surrounding Ser40 of human tyrosine hydroxylase. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from PC12 cells, untreated or forskolin-treated (30 µM), using Phospho-Tyrosine Hydroxylase (Ser40) Antibody (upper) or Tyrosine Hydroxylase Antibody #2792 (lower).

IF-IC

IF-IC

Confocal immunofluorescent images of PC-12 cells, untreated (A,C) and calf intestinal phosphatase treated (B), labeled with Phospho-Tyrosine Hydroxylase (Ser40) Antibody #2791 (green, top) and Tyrosine Hydroxylase Antibody #2792 (green, bottom) compared to an isotype control (D). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

Tyrosine hydroxylase (TH) catalyzes the rate-limiting step in the synthesis of the neurotransmitter dopamine and other catecholamines. TH functions as a tetramer, with each subunit composed of a regulatory and catalytic domain, and exists in several different isoforms (1,2). This enzyme is required for embryonic development since TH knockout mice die before or at birth (3). Levels of transcription, translation and posttranslational modification regulate TH activity. The amino-terminal regulatory domain contains three serine residues: Ser9, Ser31 and Ser40. Phosphorylation at Ser40 by PKA positively regulates the catalytic activity of TH (4-6). Phosphorylation at Ser31 by CDK5 also increases the catalytic activity of TH through stabilization of TH protein levels (7-9).

  1. Kumer, S.C. and Vrana, K.E. (1996) J Neurochem 67, 443-62.
  2. Bodeau-Péan, S. et al. (1999) J Biol Chem 274, 3469-75.
  3. Kobayashi, K. et al. (1995) J Biol Chem 270, 27235-43.
  4. Lew, J.Y. et al. (1999) Mol Pharmacol 55, 202-9.
  5. Vié, A. et al. (1999) J Biol Chem 274, 16788-95.
  6. Lindgren, N. et al. (2000) J Neurochem 74, 2470-7.
  7. Moy, L.Y. and Tsai, L.H. (2004) J Biol Chem 279, 54487-93.
  8. Lehmann, I.T. et al. (2006) J Biol Chem 281, 17644-51.
  9. Saraf, A. et al. (2007) J Biol Chem 282, 573-80.

Application References

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