Anti-Doublecortin antibody was prepared from whole goat serum produced by repeated immunizations with a synthetic peptide corresponding to a near C-terminal portion of human doublecortin conjugated to Keyhole Limpet Hemocyanin (KLH).
Konjugation:
Unconjugated
Alternative Synonym:
Goat Anti-Doublecortin Antibody, Lissencephalin-X, Doublecortex, Doublin, Lis-X, DBCN, LISX, Doublecortex Lissencephaly X-Linked (Doublecortin), Neuronal Migration Protein Doublecortin, SCLH, XLIS, DC
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Formulierung:
Liquid (sterile filtered)
Target-Kategorie:
Human
Antibody Type:
Primary Antibody
Application Verdünnung:
ELISA: 1:10,000-1:50,000, IHC: 1:200, IF Microscopy: 15 µg/mL, WB: 1:1000
Anwendungsbeschreibung:
Anti-Doublecortin Antibody has been tested in WB, IF, and IHC. Expect bands ~40.6 kDa in western blot using appropriate lysates. Positive control used: PND2-6 brain cells in western blot, PND1 in immunofluorescence, and mouse dentate gyrus for immunohist
Immunofluorescence microscopy of Anti-Doublecortin in PND1 cells. Post-natal rat pup (PND1) heterogeneous brain cells were fixed with 4% PFA, permeabilization 0.3% Triton X-100. Incubated with 15µg/mL of anti-Doublecortin primary antibody at 4C overnight. Counterstain Donkey anti-Goat IgG Dylight(TM)488 Conjugated antibody (p/n 605-741-125) at 5µg/mL for 1hr at room temperature. Nuclei were counterstained with DAPI present in mounting medium. Predicted cell localization is cytoplasm, cell projection. Note: localizes at neurite tips. Image A). DAPI only. B). Anti-Doublecortin Antibody + secondary. C). Merged A+B. D). Secondary only.
Immunofluorescence microscopy of Anti-Doublecortin in PND1 cells. Post-natal rat pup (PND1) heterogeneous brain cells were fixed with 4% PFA, permeabilization 0.3% Triton X-100. Incubated with 15µg/mL of anti-Doublecortin primary antibody at 4C overnight. Counterstain Donkey anti-Goat IgG Dylight(TM)488 Conjugated antibody (p/n 605-741-125) at 5µg/mL for 1hr at room temperature. Nuclei were counterstained with DAPI present in mounting medium. Predicted cell localization is cytoplasm, cell projection. Image A). DAPI only. B). Anti-Doublecortin Antibody + secondary. C). Merged A+B. D). Secondary only.
Immunohistochemistry of Goat Anti-Doublecortin antibody in mouse brain. Tissue: mouse dentate gyrus tissue. Antigen retrieval: heat induced (HIER) using Citrate Buffer for 20min. Primary antibody: Doublecortin antibody at 1:200 for 30min at RT. Secondary Antibody: Donkey Anti-Goat-HRP Antibody at 4µl/mL for 20min at RT. Counter Stain: Hematoxylin. Localization: Doublecortin (DCX) showed selective staining of soma and nuclei of cells in the dentate gyrus in mouse brain.
Western Blot of Goat Anti-Doublecortin Antibody. Lane 1: Opal Pre-stained MW ladder (p/n MB-210-0500). Lane 2: HEK293T Lysate (p/n W09-001-GX5) [15µg]. Lane 3: Rat brain, post natal, Whole Cell Lysate (p/n W12-001-MQ6) [15µg]. Lane 4: Rat pup PND2-6 - minimal cortex brain lysate [15µg]. Lane 5: Rat pup PND2-6 - minimal cortex brain lysate [35µg]. Primary antibody: Doublecortin antibody at 1:1000 for overnight at 4C. Secondary antibody: Donkey anti-goat secondary HRP antibody (p/n 605-703-125) at 1:40,000 for 1 hr at RT. Blocking: BlockOut (p/n MB-073) for 30 min at RT. Expected: ~40kDa. Observed size: ~45kDa.
Western Blot of Goat Anti-Doublecortin Antibody. Lane 1: Opal Pre-stained MW ladder (p/n MB-210-0500). Lane 2: Doublecortin (human) Over Expressing HEK293T Lysate C-MYC/DDK tagged. Lane 3: HEK293T Lysate (p/n W09-001-GX5). Lane 4: Ms adult brain (normal tissue) (p/n W10-000-T004). Lane 5: Rat adult brain (normal tissue) (p/n W12-000-T077). Load: 10 µg per lane. Primary antibody: Doublecortin antibody at 1:1000 for overnight at 4C. Secondary antibody: Donkey anti-goat secondary HRP antibody (p/n 605-703-125) at 1:40,000 for 1 hr at RT. Blocking: BlockOut (p/n MB-073) for 30 min at RT. Expected: ~48kDa (overexpressing). Observed size: ~48/35kDa for Doublecortin.
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