Product datasheet
Anti-Doublecortin (phospho S28) antibody - Neuronal
Marker ab23544
3 Abreviews 1 References 2 Images
Overview
Product name
Anti-Doublecortin (phospho S28) antibody - Neuronal Marker
Description
Rabbit polyclonal to Doublecortin (phospho S28) - Neuronal Marker
Tested applications
IHC-FoFr, WB, ICC/IF
Species reactivity
Reacts with: Mouse, Rat
Predicted to work with: Human
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Mouse
Doublecortin, phosphorylated at S28. Read Abcam's proprietary immunogen policy (Peptide
available as ab31511.)
Positive control
Mouse Adult Brain Whole Tissue Lysate and Mouse (0-days old) Brain Whole Tissue Lysate.
Rat adult brain tissue.
General notes
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
Properties
Form
Liquid
Storage instructions
Shipped 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.
Storage buffer
Preservative: 0.016% Sodium Azide
Constituents: 0.8% BSA, 80mM Boric Acid, 60mM Sodium chloride, 4mM EDTA, 20mM
Sodium borate, pH 7.4
Purity
Immunogen affinity purified
Primary antibody notes
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
Clonality
Polyclonal
1
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab23544 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
Notes
IHC-FoFr
1/1000.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa
(predicted molecular weight: 50 kDa).
ICC/IF
Application notes
Use at an assay dependent concentration. PubMed: 23226105
Is unsuitable for IHC-P.
Target
Function
Seems 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 iondependent signal transduction pathway. May be part with LIS-1 of an overlapping, but distinct,
signaling pathways that promote neuronal migration.
Tissue specificity
Highly 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 disease
Defects 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).
Sequence similarities
Contains 2 doublecortin domains.
Cellular localization
Cytoplasm.
Anti-Doublecortin (phospho S28) antibody - Neuronal Marker images
2
All lanes : Anti-Doublecortin (phospho S28)
antibody - Neuronal Marker (ab23544) at 1
µg/ml
Lane 1 : Mouse Adult Brain Whole Tissue
Lysate
Lane 2 : Brain (Mouse) Tissue Lysate - 0
days old (ab7188)
Lane 3 : Mouse Adult Brain Whole Tissue
Lysate with Mouse Doublecortin (phospho
S28) peptide (ab31511) at 1 µg/ml
Western blot - Doublecortin (phospho S28)
antibody - Neuronal Marker (ab23544)
Lane 4 : Brain (Mouse) Tissue Lysate - 0
days old (ab7188) with Mouse Doublecortin
(phospho S28) peptide (ab31511) at 1 µg/ml
Lane 5 : Mouse Adult Brain Whole Tissue
Lysate with Mouse Doublecortin peptide
(ab31512) at 1 µg/ml
Lane 6 : Brain (Mouse) Tissue Lysate - 0
days old (ab7188) with Mouse Doublecortin
peptide (ab31512) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor®
680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 50 kDa
Observed band size : 50 kDa
Additional bands at : 35 kDa (possible
degradation product).
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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
Immunohistochemistry - Free Floating -
post-fixed overnight in the same fixative, and
Doublecortin (phospho S28) antibody - Neuronal
cryoprotected in 20% sucrose and frozen in
Marker (ab23544)
OCT. 30µm coronal sections were cut by
S Pezet, Univ London Kings Coll, UK
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.
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