Alternative NamesDoublin, Lissencephalin-X, DBCN and Lis-X
Application(s)ICC, WB
Antibody HostMouse
Antibody TypeMonoclonal
SpecificityThe antibody reacts with two bands at ~45 kDa and ~35 kDa which shows that the mouse anti-DCX antibody binds to an epitope in the region of DCX shared by Lis-A, and Lis-B, Lis-C and Lis-D, the C terminal 360 amino acids of Lis-A. It has also been used successfully for immunocytochemistry and is an excellent marker for developing neurons.
Species ReactivityBovine, Human, Mouse, Rat
Immunogen DescriptionFull length recombinant human Lis-A isoform of Doublecortin purified from E. coli.
Application DetailsImmunocytochemistry (ICC) and Western Blotting (WB). A dilution of 1:500-1:2,000 is recommended for WB. A dilution of 1:500-1:1,000 is recommended for ICC. The optimal dilution should be determined by the end user.
TargetDoublecortin (DCX)
SpecificityThe antibody reacts with two bands at ~45 kDa and ~35 kDa which shows that the mouse anti-DCX antibody binds to an epitope in the region of DCX shared by Lis-A, and Lis-B, Lis-C and Lis-D, the C terminal 360 amino acids of Lis-A. It has also been used successfully for immunocytochemistry and is an excellent marker for developing neurons.
Target Host SpeciesHuman
Species ReactivityBovine, Human, Mouse, Rat
Antibody HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgG2a
Clone Name3E1
ConjugateUnconjugated
Immunogen DescriptionFull length recombinant human Lis-A isoform of Doublecortin purified from E. coli.
Purity DescriptionProtein G purified
FormatLyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, and sodium azide
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute with 100 µL sterile-filtered, ultrapure water to achieve a 1 mg/mL concentration. Centrifuge to remove any insoluble material.
Storage InstructionsAfter reconstitution of lyophilized antibody, aliquot and store at -20°C for a higher stability. Avoid freeze-thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Alternative NamesDoublin, Lissencephalin-X, DBCN and Lis-X
Scientific BackgroundDoublecortin (DCX, also known as Doublin, Lissencephalin-X, DBCN and Lis-X) was originally discovered since defects in the gene encoding it are causative of X-linked lissencephaly, a rare group of brain malformations resulting in a smooth cerebral cortex caused by aberrant neuronal migration during development (1,2). The name Doublecortin comes from the unusual layering of the cortex in this form of lissencephaly, which appears to have a second deep cortical layer of neurons. This layer consists of neurons which did not migrate from the subventricular zone to the normal cortical layer. Patients with this defect suffer from seizures and mental retardation. Four proteins encoded by the DCX produce bands of about 35 kDa and 45 kDa on Western blots. The 45 kDa form is known as Lis-XA while the smaller forms are generated by alternate transcription, are all missing the first 81 amino acids of Lis-XA, and are referred to as Lis-XB, Lis-XC, Lis-XD. There are minor amino acid sequence differences between these three smaller isoforms. All of these proteins contain two so-called Doublecortin domains, each about 90 amino acids long, which are believed to function in binding to microtubules, a C-terminal serine and proline rich region which may become phosphorylated in vivo. DCX is expressed very early in neuronal development, as neuroblasts become post-mitotic, but is lost as neurons mature. Developing neurons start to lose DCX expression about the time that they begin to express NeuN. Antibodies to DCX can be used to see if neurogenesis is taking place.
Rat brain extract from a postnatal 3 day animal. Two bands at ~45 kDa and ~35 kDa show that the mouse anti-DCX antibody binds to an epitope in the region of DCX shared by Lis-A, and Lis-B, Lis-C and Lis-D, the C terminal 360 amino acids of Lis-A.
Rat brain neural cultures stained with mouse anti-DCX (green), chicken polyclonal antibody to MAP2 (Catalog Number C-1382-50) (red) and DNA (blue). The mouse anti-DCX antibody (Catalog Number M-1648-100) reveals strong cytoplasmic staining in a population of small developing neurons. These cells are often found in small clumps as in these cultures, as in this example. Note that they are not positive for MAP2, which is characteristic of more mature neurons, and that the mature neuron shown is negative for DCX. This DCX antibody is therefore an excellent marker of developing neuronal cells.
Left: Analysis of doublecortin expression in cortical neuron-glial cell culture from E20 rat by Immunocytochemistry. Cells were stained with mouse antibody to doublecortin (red, 1:1,000) and chicken anti-MAP2 antibody (C-1382-50, green, 1:10,000). Blue: DAPI nuclear stain. The doublecortin antibody reveals strong cytoplasmic staining in a population of small developing neurons and their processes, while the MAP2 antibody stains dendrites and perikarya of mature neurons. Doublecortin antibody is an excellent marker of early developing neuronal cells. Right: Western blot analysis of doublecortin expression rat whole brain (green, 1:1,000). [1] protein standard, [2] adult brain, [3] embryonic E20 brain. Strong bands at 40 kDa and 45 kDa correspond to the doublecortin protein, detected only in the developing brain.
General Referencesdes Portes V et al. A novel CNS gene required for neuronal migration and involved in X-linked subcortical laminar heterotopia and lissencephaly syndrome. Cell 92:51-61 (1998). Gleeson JG, Allen KM, Fox JW, Lamperti ED, Berkovic S, Scheffer I, Cooper EC, Dobyns WB, Minnerath SR, Ross ME and Walsh CA. Doublecortin, a brain-specific gene mutated in human X-linked lissencephaly and double cortex syndrome, encodes a putative signaling protein. Cell 92:63-72 1998. Jin J, Suzuki H, Hirai S, Mikoshiba K and Ohshima T. JNK phosphorylates Ser332 of doublecortin and regulates its function in neurite extension and neuronal migration. Dev Neurobiol. 70:929-42 2010.
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