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PE Mouse anti-Doublecortin
PE Mouse anti-Doublecortin
LEFT: Analysis of doublecortin (DCX) in human embryonic stem (ES) cell-derived neurons. A sorted population of neural stem cells (NSC), that had been derived from H9 cells (WiCell, Madison, WI), were differentiated in NSC differentiation medium [containing N2 and B-27 supplements (Life Technologies), recombinant human BDNF and GDNF (Peprotech), dibutryl cyclic AMP (Sigma)] for 4 weeks. The neurons were harvested, fixed in BD Cytofix™ buffer (Cat. No. 554655), permeabilized with BD™ Phosflow Perm/Wash buffer I (Cat. No. 557885) and stained with matching concentrations of a PE Mouse IgG1, κ isotype control (shaded histogram, Cat. No. 554680) or the PE Mouse anti-Doublecortin antibody (open histogram). Histograms were derived from gated events based on light scattering characteristics for the H9-derived neurons. The dimmer peak consists of NSC and glial progenitors, and the brighter peak consists of the immature neuron population that is staining positive for doublecortin. RIGHT: Analysis of doublecortin (DCX) on human ES cell-derived neurons and glia. NSC derived from H9 cells (WiCell, Madison, WI) were differentiated in NSC differentiation medium for 11 days followed by AGM™ Astrocyte Growth Medium (Lonza) for 16 days. The neurons and glia were harvested, fixed and permeabilized, and stained with PE Mouse Anti-Doublecortin and Alexa Fluor® 647 Mouse anti-GFAP (Cat. No. 561470) monoclonal antibodies. The dot plot was derived from gated events based on light scattering characteristics for the H9-derived constituents. All flow cytometry was performed on a BD LSR™ II flow cytometry system.
LEFT: Analysis of doublecortin (DCX) in human embryonic stem (ES) cell-derived neurons. A sorted population of neural stem cells (NSC), that had been derived from H9 cells (WiCell, Madison, WI), were differentiated in NSC differentiation medium [containing N2 and B-27 supplements (Life Technologies), recombinant human BDNF and GDNF (Peprotech), dibutryl cyclic AMP (Sigma)] for 4 weeks. The neurons were harvested, fixed in BD Cytofix™ buffer (Cat. No. 554655), permeabilized with BD™ Phosflow Perm/Wash buffer I (Cat. No. 557885) and stained with matching concentrations of a PE Mouse IgG1, κ isotype control (shaded histogram, Cat. No. 554680) or the PE Mouse anti-Doublecortin antibody (open histogram). Histograms were derived from gated events based on light scattering characteristics for the H9-derived neurons. The dimmer peak consists of NSC and glial progenitors, and the brighter peak consists of the immature neuron population that is staining positive for doublecortin. RIGHT: Analysis of doublecortin (DCX) on human ES cell-derived neurons and glia. NSC derived from H9 cells (WiCell, Madison, WI) were differentiated in NSC differentiation medium for 11 days followed by AGM™ Astrocyte Growth Medium (Lonza) for 16 days. The neurons and glia were harvested, fixed and permeabilized, and stained with PE Mouse Anti-Doublecortin and Alexa Fluor® 647 Mouse anti-GFAP (Cat. No. 561470) monoclonal antibodies. The dot plot was derived from gated events based on light scattering characteristics for the H9-derived constituents. All flow cytometry was performed on a BD LSR™ II flow cytometry system.
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BD Pharmingen™
DCX, Lissencephalin-X, Lis-X, Doublin, Doublecortex, DBCN, LISX
Human (QC Testing), Mouse (Reactivity Confirmed in Development)
Mouse IgG1
Mouse Doublecortin aa. 211-317 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_10643766
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation and Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
561505 Rev. 1
抗体の詳細
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30/Doublecortin

The cerebral cortex is composed of multiple layers of neurons of distinct types and functions, which perform in unison to orchestrate cognitive function. The formation of the cortex relies on complex signaling mechanisms, which mediate the migration of newly formed neurons from deep within the brain to the superficial regions. Defects in neuronal migration and disruption of the multi-layered cortex are apparent in conditions such as X-linked lissencephaly (XLIS) and subcortical laminar heterotopia (SCLH) or "double cortex" (DC) syndrome. Mutations in the doublecortin gene have been linked to these brain disorders. Doublecortin is highly expressed in developing brain, primarily in migrating neurons. It is significantly homologous with the N-terminal region of the product of the KIAA0369 gene, a protein that is also similar to the CaM kinase family in its C-terminal region. Doublecortin has four potential MAP kinase family phosphorylation sites and a putative site for Abl tyrosine phosphorylation. Thus, doublecortin is thought to be an integral component of tyrosine kinase signal transduction pathways that regulate neuronal migration and development of the cerebral cortex.

The 30/Doublecortin monoclonal antibody reacts with mouse and human doublecortin.

561505 Rev. 1
フォーマットの詳細
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
561505 Rev.1
引用&参考文献
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Development References (3)

  1. Gleeson JG, Allen KM, Fox JW, et al. Doublecortin, a brain-specific gene mutated in human X-linked lissencephaly and double cortex syndrome, encodes a putative signaling protein. Cell. 1998; 92(1):63-72. (Biology). View Reference
  2. Magavi SS, Leavitt BR, Macklis JD. Induction of neurogenesis in the neocortex of adult mice. Nature. 2000; 405(6789):951-955. (Biology). View Reference
  3. des Portes V, Pinard JM, Billuart P, et al. A novel CNS gene required for neuronal migration and involved in X-linked subcortical laminar heterotopia and lissencephaly syndrome. Cell. 1998; 92(1):51-61. (Biology). View Reference
561505 Rev. 1

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.