Anti-Doublecortin抗体
参阅全部 Doublecortin 一抗
兔多克隆抗体to Doublecortin
Rabbit
Please note: Low dilutions of this antibody can cause high background in IHC. Please use as high a dilution as possible. Optimal working dilutions are batch dependent.
适用于: WB, IHC-FoFr, ICC/IF, IHC-Fr, IHC-Pmore details
与反应: Mouse, Rat, Quail
预测可用于: Human, Cynomolgus monkey
Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Human Doublecortin.
参阅Abcam的专有抗源政策 (Peptide available as ab19804.)
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Liquid
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.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab18723于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | (6) | Use a concentration of 1 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 40-45 kDa). |
| IHC-FoFr | (11) | 1/1300 - 1/2000. |
| ICC/IF | (7) | Use a concentration of 1 - 5 µg/ml. |
| IHC-Fr | (7) | Use a concentration of 1 µg/ml. This antibody works well on Formalin fixed frozen sections, but will not work on fresh-frozen sections. For IHC-Fr, we recommend to use PFA perfusion fixed tissues. After dissection, fix the tissue further in PFA overnight, wash in PBS and incubate overnight in 30% sucrose/PBS, and then embed in OCT compound and cryosection. Read More |
| IHC-P | (16) | Use a concentration of 0.05 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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Unigene: 12871 Mouse
Unigene: 121471 Rat
Entrez Gene: 1641 Human
Entrez Gene: 13193 Mouse
Omim: 300121 Human
SwissProt: O43602 Human
SwissProt: O88809 Mouse
Unigene: 34780 Human
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Lis X antibody
Lis-X antibody
Lissencephalin X antibody
Lissencephalin-X antibody
Lissencephaly X linked antibody
Lissencephaly X linked doublecortin antibody
LISX antibody
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OTTHUMP00000023859 antibody
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DC antibody
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Doublecortin antibody (ab18723)
Immunohistochemical analysis of formalin-fixed paraffin-embedded mouse brain labelling doublecortin with ab18723 at a dilution of 0.8µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab18723 anti doublecortin antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Immunocytochemistry/ Immunofluorescence - Anti-Doublecortin antibody (ab18723)
ab18723 staining Doublecortin in Rt Primary Neurons DIV1 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab18723 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Doublecortin antibody (ab18723)
IHC image of Doublecortin staining in a section of frozen PFA perfusion fixed 6 week old rat dentate gyrus. After dissection, the tissue was further fixed in PFA overnight, washed in PBS and then incubated overnight in 30% sucrose. It was then embedded in OCT and cryosectioned. Non-specific protein-protein interactions were blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M glycine and 1% (w/v) BSA for 1h at room temperature. The section was incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab18723 at 1/1300 dilution and then incubated with ab150080 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preabsorbed, (Shown in red) 1/1000) for 1 hour at room temperature. DAPI was used to stain the cell nuclei (blue). The secondary-only control insert image is taken from an identical assay without primary antibody.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
For IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Doublecortin antibody (ab18723)
IHC image of Doublecortin staining in a section of frozen PFA perfusion fixed 8 week old mouse SVZ. After dissection, the tissue was further fixed in PFA overnight, washed in PBS and then incubated overnight in 30% sucrose. It was then embedded in OCT and cryosectioned. Non-specific protein-protein interactions were blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M glycine and 1% (w/v) BSA for 1h at room temperature. The section was incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab18723 at 1/2000 dilution and then incubated with ab150080 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preabsorbed, (Shown in red) 1/1000) for 1 hour at room temperature. DAPI was used to stain the cell nuclei (blue). The secondary-only control insert image is taken from an identical assay without primary antibody.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
For IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Doublecortin antibody (ab18723)
IHC image of Doublecortin staining in a section of frozen PFA perfusion fixed 8 week old mouse dentate gyrus. After dissection, the tissue was further fixed in PFA overnight, washed in PBS and then incubated overnight in 30% sucrose. It was then embedded in OCT and cryosectioned. Non-specific protein-protein interactions were blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M glycine and 1% (w/v) BSA for 1h at room temperature. The section was incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab18723 at 1/2000 dilution and then incubated with ab150080 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preabsorbed, (Shown in red) 1/1000) for 1 hour at room temperature. DAPI was used to stain the cell nuclei (blue). The secondary-only control insert image is taken from an identical assay without primary antibody.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
For IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.
Western blot - Anti-Doublecortin antibody (ab18723)
All lanes : Anti-Doublecortin antibody (ab18723) at 1 µg/ml
Lane 1 : Mouse brain tissue lysate - total protein (0 days)
Lane 2 : Rat brain tissue lysate - total protein (0 days)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 40-45 kDa
Observed band size: 45 kDawhy is the actual band size different from the predicted?
Blocking buffer: 3% Milk
Gel type: MOPS
Exposure Time: 1 minute 30 seconds
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Doublecortin antibody (ab18723)This image is courtesy of an anonymous abreview.
IHC-FoFr image of Doublecortin staining in Mouse adult dentate gyrus sections using ab18723 (1/100 dilution). The sections were fixed in paraformaldehyde and permeabilized using 1x TBST. The sections were then blocked using 10% donkey serum for 1 hour at 25°C. ab18723 was diluted 1/100 and incubated with the sections for 12 hours at 25°C. The secondary antibody used was Donkey anti-rabbit conjugated to Cy3 Dye (1/500 dilution). DAPI was used to stain the nuclei.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Doublecortin antibody (ab18723)
IHC image of ab18723 staining in Mouse 8 weeks brain formalin fixed paraffin embedded tissue section, performed on a Leica BONDTM system using the standard protocol F (with no post primary). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18723, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. Secondary-only control image is shown as insert.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-Doublecortin antibody (ab18723)
Anti-Doublecortin antibody (ab18723) at 1 µg/ml + Mouse brain tissue lysate - total protein (0 days) (ab7188) at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 40-45 kDa
Observed band size: 45 kDawhy is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Doublecortin antibody (ab18723)
IHC image of ab18723 staining in rat 6 week brain formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) for 20 mins. The section was then incubated with ab18723, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Doublecortin antibody (ab18723)This image is courtesy of Francois Guillemot, NIMR, UK
Doublecortin expression in the dentate gyrus of a 1 month-old mouse brain. Doublecortin staining using ab18723 (1/500) in the dentate gyrus of a 1 month-old mouse brain. The mouse has been perfused with paraformaldehyde 4% (50ml). After dissection, the brain has been incubated overnight in sucrose 20%, embedded in OCT and cryosectioned (10 µm). No antigen retrieval was used. The secondary antibody used was a non-Abcam Goat anti-rabbit Alexa488.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Doublecortin antibody (ab18723)
ab18723 staining 6 week rat brain tissue dentate gyrus (DG) by IHC-P using rabbit-specific EXPOSE IHC detection kit (ab80437). Formalin fixed paraffin embedded tissue sections were pre-treated using heat mediated antigen retrieval (using a pressure cooker) with sodium citrate buffer (pH6) for 30 mins. The section was incubated with ab18723, 0.1µg/ml, for 1 hour at room temperature. DAB was used as the chromogen and the section was counterstained with haematoxylin and mounted with DPX.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Doublecortin antibody (ab18723)This image is courtesy of Carl Hobbs, King's College London, United Kingdom
ab18723 at 1/200 staining mouse E18 body T/S spinal cord tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retireval step was perfomed before incubation with the antibody for 24 hours. A biotinylated goat antibody was used as the secondary.
Immunocytochemistry/ Immunofluorescence - Anti-Doublecortin antibody (ab18723)This image is courtesy of Randal Moldrich, CNRS UMR7637, ESPCI, France
Doublecortin antibody (ab18723; green) labeling cell extensions from rat embryos consistent with dendrite morphology in 4 day old cultures. Preincubation of ab18723 with its immunising peptide (ab19804) quenched immunostaining (see review).
Dorsal root ganglion explants were dissected from 16 day-old rat embryos and cultured for 4 days in vitro with Neurobasal Medium containing 10% fetal calf serum and B27 supplement. Immunocytochemistry: All steps were performed in PBS. Cells or explants were fixed in 4% PFA for 15min, permeabilised with 0.1% TX100 for 10min and blocked with 5% BSA, 0.1% TX100 for 45min. ab18723 was incubated at 5µg/ml for 12h in 5% BSA, 0.1% TX100 at 4°C. For peptide blocking experiments preincubation of the peptide (ab19804; 250µg/ml) and antibody (5µg/ml ) was performed for 2h at 37°C. Cultures were washed (3x) of primary antibody solution. Goat anti-rabbit AlexaFluor 488 was used as secondary antibody (1/400) in 5% BSA, 0.1% TX100 for 2h at R
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Doublecortin antibody (ab18723)This image is courtesy of Carl Hobbs, King's College London, United Kingdom
Immunohistochemistical staining (on formaldehyde/PFA-fixed paraffin-embedded sections) of Doublecortin antibody - Neuronal Marker (ab18723) on Quail Tissue sections (E6/7 brain (Saggital section). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Primary Antibody ab18723 incubated at 1/400 for 2 hours RT. Secondary Antibody: Biotin conjugated goat anti rabbit Ig (1/300).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Doublecortin antibody (ab18723)This image is courtesy of Carl Hobbs, King's College London, United Kingdom
ab18723 at 1/2000 staining mouse brain svr: progenitor olfactory neurones by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step was performed before incubation with the antibody for 16 hours. An Alexa-Fluor ® 488 conjugated goat antibody was used as the secondary (green). The tissue was also stained for Ki67 (shown in red).
The MIP image was derived from Apotome-generated Z-stacks from the greyscale image of each of the channels in the MIP.
