Mouse Adult Brain Whole Tissue Lysate and Mouse (0-days old) Brain Whole Tissue Lysate. Rat adult brain tissue.
Doublecortin (DCX) undergoes a cycle of dephosphorylation and phosphorylation that regulates its binding to microtubules in growing neurites. The N-terminus serine-28 is a major site of phosphorylation for DCX; this was identified by Graham et al (2004). Single mutation of Ser-28 reduced but did not abolish phosphorylation; DCX is subject to complex multi-site phosphorylation
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Primary antibody notesDoublecortin (DCX) undergoes a cycle of dephosphorylation and phosphorylation that regulates its binding to microtubules in growing neurites. The N-terminus serine-28 is a major site of phosphorylation for DCX; this was identified by Graham et al (2004). Single mutation of Ser-28 reduced but did not abolish phosphorylation; DCX is subject to complex multi-site phosphorylation
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
Use at an assay dependent concentration. PubMed: 23226105
Application notesIs unsuitable for IHC-P.
FunctionSeems to be required for initial steps of neuronal dispersion and cortex lamination during cerebral cortex development. May act by competing with the putative neuronal protein kinase DCAMKL1 in binding to a target protein. May in that way participate in a signaling pathway that is crucial for neuronal interaction before and during migration, possibly as part of a calcium ion-dependent signal transduction pathway. May be part with LIS-1 of an overlapping, but distinct, signaling pathways that promote neuronal migration.
Tissue specificityHighly expressed in neuronal cells of fetal brain (in the majority of cells of the cortical plate, intermediate zone and ventricular zone), but not expressed in other fetal tissues. In the adult, highly expressed in the brain frontal lobe, but very low expression in other regions of brain, and not detected in heart, placenta, lung, liver, skeletal muscles, kidney and pancreas.
Involvement in diseaseDefects in DCX are the cause of lissencephaly X-linked type 1 (LISX1) [MIM:300067]; also called X-LIS or LIS. LISX1 is a classic lissencephaly characterized by mental retardation and seizures that are more severe in male patients. Affected boys show an abnormally thick cortex with absent or severely reduced gyri. Clinical manifestations include feeding problems, abnormal muscular tone, seizures and severe to profound psychomotor retardation. Female patients display a less severe phenotype referred to as 'doublecortex'. Defects in DCX are the cause of subcortical band heterotopia X-linked (SBHX) [MIM:300067]; also known as double cortex or subcortical laminar heterotopia (SCLH). SBHX is a mild brain malformation of the lissencephaly spectrum. It is characterized by bilateral and symmetric plates or bands of gray matter found in the central white matter between the cortex and cerebral ventricles, cerebral convolutions usually appearing normal. Note=A chromosomal aberration involving DCX is found in lissencephaly. Translocation t(X;2)(q22.3;p25.1).
Immunohistochemistry free floating - Anti-Doublecortin (phospho S28) antibody - Neuronal Marker (ab23544)This image is courtesy of Sophie Pezet, King's College London, United Kingdom
Immunofluorescent staining for Doublecortin (phospho S28) in the rat habenular nucleus using Rabbit polyclonal to Doublecortin (phospho S28). The staining pattern observed with ab23544 appears to be specific, however, the staining is not very strong or contrasted - this might be expected for detection of a modification protein in unstimulated tissue.
Protocol details: Rats were intracardially perfused with 4% paraformaldehyde. Whole brain tissue was post-fixed overnight in the same fixative, and cryoprotected in 20% sucrose and frozen in OCT. 30µm coronal sections were cut by cyrostat for use in free floating IHC. Primary antibody ab123544 was incubated overnight at 1/1000 at room temperature. Secondary antibody Alexa fluor 488 1/1000 was incubated for 2 hours at room temperature.
References for Anti-Doublecortin (phospho S28) antibody - Neuronal Marker (ab23544)
This product has been referenced in:
Feltrin D et al. Growth cone MKK7 mRNA targeting regulates MAP1b-dependent microtubule bundling to control neurite elongation. PLoS Biol10:e1001439 (2012).
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