Floating sections were rinsed 3 × 5 min in PBS and incubated for 1 h at room temperature in PBS containing 1% bovine serum albumin and 0.1% Triton-X-100, followed by 72 h of incubation with primary antibody. For BrdU, the formamide treatment described above was also applied. The following primary antibody dilutions were used: 1:1000 mouse anti-BrdU (#AB6326, Abcam, Cambridge, UK), 1:500 mouse anti-NeuN (#MAB377, Millipore), 1:300 rabbit anti-cleaved caspase-3 (#2305-PC, Trevigen,Gaithersburg, MD, USA), 1:2000 anti-GFP (#A6455, Thermofisher, Life Technologies, Carlsbad, CA, USA) and 1:500 goat polyclonal anti-DCX (#8066, Santa Cruz, Dallas, TX, USA). Detection was with Alexa dye-conjugated antibodies (Molecular Probes, Eugene, OR, USA, Thermofisher, Waltham, MA, USA) at a concentration of 1:500. Nuclei were visualized with Hoechst-33342 (1:1000). Tiled images were acquired with a LSM-780 confocal microscope (Carl Zeiss, Oberkochen, Germany) using 10 × , 20 × and 40 × objectives. Images shown are typically from maximum projections of 5 × z-planes.
Goat polyclonal anti dcx
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Goat polyclonal anti-DCX is a laboratory reagent that binds to the doublecortin (DCX) protein. DCX is a microtubule-associated protein expressed in neuronal precursor cells and developing neurons. This product can be used to detect and study DCX expression in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 780 confocal microscope, by Zeiss (3 mentions)
Biotinylated anti mouse igg, by GE Healthcare (2 mentions)
Mouse monoclonal anti brdu, by Agilent Technologies (2 mentions)
Rabbit anti gfap, by Agilent Technologies (2 mentions)
Alexa dye conjugated antibodies, by Thermo Fisher Scientific (1 mentions)
Mouse anti neun, by Merck Group (1 mentions)
Mouse monoclonal anti neun, by Merck Group (1 mentions)
Avidin biotin complex reagent, by Vector Laboratories (1 mentions)
Mouse anti ed1, by Bio-Rad (1 mentions)
Mouse anti brdu, by BD (1 mentions)
Mouse anti brdu, by Abcam (1 mentions)
5 protocols using goat polyclonal anti dcx
1
Immunofluorescence Labeling and Microscopy
2
Neurogenesis and Cell Proliferation Assay
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Brain removal, processing, and immunohistochemical techniques to detect BrdU and doublecortin (DCX) were performed as previously described (Rabaneda et al., 2008 (link)). Antibodies used were mouse monoclonal anti-BrdU (1:100) from Dako (Hamburg, Germany), goat polyclonal anti-DCX (1:200, Santa Cruz Biotech.), AlexaFluor 488 donkey anti-mouse and AlexaFluor 594 (both at 1:1000, from Life Tech.), and biotinylated anti-mouse IgG (1:250) from GE Healthcare (Albany, NY).
3
Immunohistochemical Detection of Neural Markers
4
Hippocampal Immunohistochemistry for Astrocytes, Microglia, and Neurogenesis
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Thirty-micrometer thick coronal sections were cut through the entire hippocampus from fixed brains using a cryocut, collected serially in 24-well plates containing cryobuffer, and stored at −20°C until further analysis. Every 15th or 20th section through the entire septotemporal axis of the hippocampus were processed for immunohistochemical analysis of astrocytes, microglia, and hippocampal neurogenesis (6–8 animals/group). The immunohistochemical studies comprised detection of glial fibrillary acidic protein (GFAP) positive astrocytes, ED-1 positive activated microglia, 5-bromodeoxyuridine (BrdU) positive newly born cells, and doublecortin (DCX) positive newly born neurons. The methods employed are described in our previous reports [17 (link),45 (link),49 ,50 (link)]. The primary antibodies comprised rabbit anti-GFAP (1:1000, Dako, Santa Clara, CA), mouse anti ED-1 (1:1000, Bio-Rad, Hercules, CA), mouse anti-BrdU (1:200, BD, San Jose, CA), and goat polyclonal anti-DCX (1:250; Santa Cruz Biotech, Santa Cruz, CA). The secondary antibodies comprised anti-goat, anti-mouse, or anti-rabbit IgG (Vector Laboratories, Burlingame, CA). The avidin-biotin complex reagent and chromogen kits (vector gray or diaminobenzidine) were obtained from Vector Labs. The sections were next mounted on gelatin-coated slides, dehydrated, cleared, and coverslipped.
5
Immunohistochemistry for Hippocampal Neurogenesis
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We performed immunohistochemistry for visualizing glial fibrillary acidic protein (GFAP) positive astrocytes, BrdU+ newly born cells, and doublecortin (DCX) positive newly born neurons in the hippocampus. The methods employed for cutting thirty-micrometer thick coronal sections through the entire forebrain, collection, and storage of sections in cryobuffer, and immune-histochemical methods are described in our previous reports [15 (link), 46 (link), 52 (link), 53 (link)]. GFAP, BrdU, and DCX immunohistochemistry were done on every 15th or 20th section through the hippocampus's entire septotemporal axis in each animal. The primary antibodies comprised rabbit anti-GFAP (1:2000; Dako, Santa Clara, CA), mouse anti-BrdU (1:500; Abcam), and goat polyclonal anti-DCX (1:300; Santa Cruz, Dallas, TX). The secondary antibodies comprised anti-rabbit, anti-mouse, or anti-goat IgG’s (Vector Laboratories, Burlingame, CA). The avidin-biotin complex reagent and chromogen kits (vector gray or diaminobenzidine) were also purchased from Vector Labs. The sections were fixed on gelatin-coated slides, dehydrated, cleared, and coverslipped.
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