- 48-1200 - Invitrogen Antibodies DCX antibody

Submitted references Krüppel-like factor 7 attenuates hippocampal neuronal injury after traumatic brain injury.

Li WY, Fu XM, Wang ZD, Li ZG, Ma D, Sun P, Liu GB, Zhu XF, Wang Y
Neural regeneration research 2022 Mar;17(3):661-672

High-fat diet increases gliosis and immediate early gene expression in APOE3 mice, but not APOE4 mice.

Jones NS, Watson KQ, Rebeck GW
Journal of neuroinflammation 2021 Sep 18;18(1):214

Bio-electrosprayed human neural stem cells are viable and maintain their differentiation potential.

Helenes González C, Jayasinghe SN, Ferretti P
F1000Research 2020;9:267

Fluoxetine effects on behavior and adult hippocampal neurogenesis in female C57BL/6J mice across the estrous cycle.

Yohn CN, Shifman S, Garino A, Diethorn E, Bokka L, Ashamalla SA, Samuels BA
Psychopharmacology 2020 May;237(5):1281-1290

Increased Expression of Y-Encoded Demethylases During Differentiation of Human Male Neural Stem Cells.

Pottmeier P, Doszyn O, Peuckert C, Jazin E
Stem cells and development 2020 Dec 1;29(23):1497-1509

Social instability is an effective chronic stress paradigm for both male and female mice.

Yohn CN, Ashamalla SA, Bokka L, Gergues MM, Garino A, Samuels BA
Neuropharmacology 2019 Dec 1;160:107780

Spontaneous Differentiation of Human Neural Stem Cells on Nanodiamonds.

Taylor AC, González CH, Ferretti P, Jackman RB
Advanced biosystems 2019 Apr;3(4):e1800299

3D Culture Method for Alzheimer's Disease Modeling Reveals Interleukin-4 Rescues Aβ42-Induced Loss of Human Neural Stem Cell Plasticity.

Papadimitriou C, Celikkaya H, Cosacak MI, Mashkaryan V, Bray L, Bhattarai P, Brandt K, Hollak H, Chen X, He S, Antos CL, Lin W, Thomas AK, Dahl A, Kurth T, Friedrichs J, Zhang Y, Freudenberg U, Werner C, Kizil C
Developmental cell 2018 Jul 2;46(1):85-101.e8

Dynamic microtubule association of Doublecortin X (DCX) is regulated by its C-terminus.

Moslehi M, Ng DCH, Bogoyevitch MA
Scientific reports 2017 Jul 12;7(1):5245

Use of induced pluripotent stem cell derived neurons engineered to express BDNF for modulation of stressor related pathology.

Liu G, Rustom N, Litteljohn D, Bobyn J, Rudyk C, Anisman H, Hayley S
Frontiers in cellular neuroscience 2014;8:316

Interaction between nonviral reprogrammed fibroblast stem cells and trophic factors for brain repair.

Liu G, Anisman H, Bobyn J, Hayley S
Molecular neurobiology 2014 Oct;50(2):673-84

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell and tissue extracts (30 µg lysate) of SH-SY5Y (Lane 1) and Rat Brain (Lane 2) respectively. The blots were probed with Anti-Doublecortin/DCX Rabbit polyclonal Antibody (Product # 48-1200, 2 µg/mL) and detected by chemiluminescence Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution). Isoforms of 41 kDa and 49 kDa corresponding to Doublecortin/DCX was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0341BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Doublecortin was achieved by transfecting SH-SY5Y cells with Doublecortin specific siRNAs (Silencer® select Product # s3976, s3978). Western blot analysis (Fig. a) was performed using whole cell extracts from the Doublecortin knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The bot was probed with Doublecortin Polyclonal Antibody (Product # 48-1200, 1µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Doublecortin. An uncharacterized band (*) was also observed at ~70kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Doublecortin was performed using 70% confluent log phase U-87MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Doublecortin/DCX Rabbit Polyclonal Antibody (Product # 48-1200) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Doublecortin was done on SH-SY5Y cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Doublecortin Rabbit Polyclonal Antibody (481200, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIGURE 3 In vitro iPSCs and neural progenitor cells produced from adult mouse fibroblasts (see Materials and Methods). The panels show that control (absence of the 20866 vector) tail cells still retain their original shape on Day 47 after culturing (A) . The presence of the mOrange marker suggests that the fibroblasts have successfully integrated the 20866 plasmid (B) . The iPSCs were further characterized by an alkaline phosphatase live stain (C) , as well as staining of the three cell-surface stage-specific embryonic antigens: SSEA-1 to -4 (D-F) , TRA-1-60 (G) , TRA-1-81 (H) , and Oct-4 (I) . By Day 18, iPSCs began displaying projections (J) and further differentiation was clearly evident by Days 20, 23, and 28 (K-M) , respectively. By the later time point (M) , the cells had morphology consistent with that of neural progenitor cells and displayed a robust induction of the immature neuronal marker, doublecortin (N) .

Supportive validation

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Experimental details: Fluorescently stained images of neurons, which have been differentiated from hNSCs after a) 25 DIV and b) NeuN (green), DCX (red) and Hoechst (blue). All scale bars 100 um.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 FRAP analysis reveals that GFP-DCX dynamics can be influenced by agents that alter MT stability. COS-1 cells were transfected to express ( A ) GFP-DCX WT or ( C ) GFP-DCX DeltaC. Cells were exposed to nocodazole (20 uM, 2 h) to depolymerise MTs or to taxol (10 uM, 1 h) to stabilize MT polymers. A small ROI (indicated by white rectangle in each cell image) was photobleached in ( A ) and ( C ) and the fluorescence recovery was subsequently monitored post-bleach at 3 s intervals for 60 s. Plots of the recovery of fluorescence in the small area of bleach for ( B ) DCX WT and ( D ) DCX DeltaC constructs before and after nocodazole or taxol treatments are shown. Regression values for the accuracy of each curve fit are indicated. The insets represent the initial recovery of fluorescence in the photobleached area for first three time-points (0-6 s of post-bleach) with the line of best fit used for the calculation of the initial recovery rates. ( E ) Equivalent expression of the GFP-DCX constructs was detected in the presence and absence of nocodazole or taxol by immunoblotting for DCX (upper panel), with equivalent protein loading detected by immunoblotting for total alpha-tubulin (lower panel). Scale bars represent 10 um. See Fig. 4 for quantitative pooled data.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 7 HFD does not alter neurogenesis or spine density in APOE 3 and APOE 4 mice. A Image of spines quantified from Golgi staining. B Image of DCX positive cells. C - D Quantification of spine density on the Apical Oblique and Basal Shaft. E Quantification of DCX positive cells

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 10 KLF7 increases neural proliferation following TBI. (A-C) Immunofluorescence staining of BrdU (proliferating neurons, red, stained by Alexa 555), DCX (newly generated neurons, red, stained by Alexa 555), and GFAP (astrocytes, green, stained by fluorescein isothiocyanate) with nuclear staining (DAPI; blue) in the ipsilateral hippocampus dentate gyrus (DG) in the three groups at 3 and 7 days following TBI. Compared with the TBI + AAV-NC group, BrdU-positive cells increased in the TBI + AAV-KLF7 group at 3 and 7 days following TBI. DCX-positive cells exhibited a similar pattern of change under the different treatment conditions. Scale bars: 100 mum in A, 200 mum in B and C. (D, E) Quantification of the number of BrdU-positive cells in the DG at 3 (D) and 7 (E) days following TBI. (F) Quantification of the density of DCX-positive cells at 3 days following TBI. Data are expressed as the mean +- SD ( n = 4). * P < 0.05, *** P < 0.001 (one-way analysis of variance followed by Tukey's post hoc test). AAV: Adeno-associated virus; BrdU: 5-bromo-2'-deoxyuridine; DAPI: 4',6-diamidino-2-phenylindole; DCX: doublecortin; GFAP: glial fibrillary acidic protein; KLF7: Kruppel-like factor 7; NC: negative control; TBI: traumatic brain injury.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 5 (A) Immature neuronal cells that were cultured in a Dopaminergic Neuronal Progenitor Medium began to show morphological signs of neuronal maturity between Day 29-50 (a,b,c) and after 6 months these cells displayed further signs of mature network formation (d). Importantly, control tail cells that were not subject to the reprogramming procedures kept their original shape at 6 months (e). As well, during this time, expression of the early immature neuronal cell marker, DCX, was greatly diminished (f), together with increased expression of the mature general neuronal cell marker, microtubule associated protein 2 (MAP2), (g), and specific dopaminergic neuronal marker, tyrosine hydroxylase (TH), (h) was evident. Further confirmation that the reprogrammed neurons were mature dopaminergic neurons was obtained using HPLC (i); wherein tail cells that were re-programmed (Tail-Dopamine cells) displayed significantly increased dopamine levels, compared to non-reprogrammed controls (Tail cells). Furthermore, dopamine levels in non-reprogrammed controls were, as expected, quite low (Tail tissue), whereas freshly dissected adult midbrain tissue had the expected highest concentration of dopamine (Brain tissue). * p < 0.05, relative to control tails. (B) Shows the time course for dopaminergic neuron derivation. The adult mouse (aged 2 months) fibroblasts were reprogrammed by vector 20866 with c-Myc, Klf4, Oct4, and Sox2 genes and an mOrange marker was used to detect gene expression a

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 4 As shown in the bar graph on the left (A) , the single icv LPS infusion (2 mug) plus a 3-week twice-daily different stressor challenge (LPS + stress) increased plasma corticosterone, relative to vehicle treatment (Veh). Icv infusion of either the iPSC-derived neural progenitor cells alone (LPS + stress + iPSC) or in combination with the cDNA BDNF (LPS + stress + iPSC + BDNF) attenuated the plasma corticosterone rise induced by the LPS + stress treatment. The photomicrograph on the right (B) displays doublecortin (DCX) immunoreactivity in the subventricular zone (SVZ) in response to vehicle (a), LPS + stressor exposure (b), LPS + stressor exposure + iPSC-derived neural progenitor cells (c), or LPS + stressor exposure + iPSC-derived neural progenitors + BDNF cDNA (d). As quantified in the bottom bar graph, the LPS + stressor exposure alone or in combination with the iPSC-derived neural progenitor cells significantly reduced the number of DCX+ cells within the SVZ ( p < 0.05). However, infusion of the iPSC derived neural progenitor cells that were transfected with cDNA BDNF completely prevented this DCX reduction. * p < 0.05 relative to Veh treatment, o p < 0.05 relative to LPS + stress treatment.

48-1200

Antibody data