Cell Signaling Technology

Product Pathways - Neuroscience

Phospho-Doublecortin (Ser334) Antibody #3453

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-F H M R Endogenous 45 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-F=Immunofluorescence (Frozen)
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-Doublecortin (Ser334) Antibody detects endogenous levels of doublecortin only when phosphorylated at Ser334.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser334 of doublecortin. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from neonatal mouse brain and fetal rat brain using Phospho-Doublecortin (Ser334) Antibody. The phospho-specificity of the antibody was verified by blocking with a phospho- or non-phosphopeptide and by treating the membrane with (+) or without (-) calf intestinal phosphatase (CIP) after western transfer.

IF-F

IF-F

Confocal immunofluorescent analysis of postnatal day 14 rat brain, untreated (left) or λ-phosphatase treated (right), using Phospho-Doublecortin (Ser334) Antibody (green) and GFAP (GA5) Mouse mAb #3670 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

Mutations in Doublecortin cause Lissencephaly (smooth brain), a neuronal migration disorder characterized by epilepsy and mental retardation (1). Doublecortin is a microtubule associated protein that stabilizes and bundles microtubules. A conserved doublecortin domain mediates the interaction with microtubules, and interestingly most missense mutations cluster in this domain (2). Kinases JNK, CDK5 and PKA phosphorylate doublecortin. JNK phosphorylates Thr321, Thr331 and Ser334 while PKA phosphorylates Ser47 and CDK5 phosphorylates Ser297 (3-5). Phosphorylation of Ser297 lowers the affinity of doublecortin to microtubules. Furthermore, mutations of Ser297 result in migration defects (5).

Doublecortin phosphorylated at Ser334 is enriched in growth cones and affects neurite outgrowth and neuronal migration (3).

  1. Gleeson, J.G. et al. (1998) Cell 92, 63-72.
  2. Reiner, O. et al. (2004) Cell Cycle 3, 747-751.
  3. Gdalyahu, A. et al. (2004) EMBO J. 23, 823-832.
  4. Schaar, B.T. et al. (2004) Neuron 41, 203-213.
  5. Tanaka, T. et al. (2004) Neuron 41, 215-227.

Application References

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