After being anesthetized with euthatal (from Merck) (60 mg/kg), the Nrg1-reporting mice (2 months old) were transcardially perfused with PBS (2 ml/g of body weight), followed by 4% PFA in PBS. Brains were harvested, incubated in 4% PFA overnight, and dehydrated at 4 °C in two steps with 20% and 30% sucrose in PBS. Brains were frozen in OCT (catalog #14-373-65; Fisher) and sectioned into 40 µm slices on a cryostat microtome (Bosch Microm HM550) at − 20 °C. Brain slices were permeabilized with 0.3% TritonX 100 and 5% BSA in PBS and incubated with primary antibodies at 4 °C overnight. The brain slices were not treated with Triton- × 100 when staining with anti-GAD67 antibodies. After washing with PBS for three times, samples were incubated with Alexa Fluor-488 or -647 secondary antibodies (1:1000, ThermoFisher Scientific) for 1 h at room temperature. Samples were mounted with Vectashield mounting medium (Vector) and images were taken by Leica TCS SP8 confocal microscope. The following primary antibodies were used: rabbit anti-NeuN (1:500, Abcam, ab177487), rabbit anti-TH (1:250, Millipore, MAB152), mouse anti-NRGN (1:200, R&D, MAB7947), mouse anti-PV (1:500, Sigma, P3088), mouse anti-GFAP (1:250, Millipore, MAB360); rabbit anti-S100β (1:200, Abcam, Ab52642) and mouse anti-GABA (1:1000, Invitrogen, PA5-32241).
Ab52642
Manufactured by Thermo Fisher Scientific
Ab52642 is a laboratory equipment product designed for general use in research and testing applications. It serves as a core component for various analytical procedures. The detailed specifications and intended use of this product are not available.
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Lab products found in correlation
Ab177487, by Merck Group (1 mentions)
Alexa fluor 488 or 647 secondary antibodies, by Thermo Fisher Scientific (1 mentions)
P3088, by Merck Group (1 mentions)
Mab360, by Abcam (1 mentions)
Tcs sp8 confocal microscope, by Leica (1 mentions)
Vectashield mounting medium, by Vector Laboratories (1 mentions)
Mab360, by Merck Group (1 mentions)
Gfp 1020, by Rockland Immunochemicals (1 mentions)
Sc 17320, by Santa Cruz Biotechnology (1 mentions)
Mab377, by Merck Group (1 mentions)
2 protocols using ab52642
1
Immunohistochemistry of Nrg1-Reporting Mouse Brain
2
Immunostaining for Neural Cell Markers
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Sections without signs of tissue damage were chosen for immunostaining (see also Figure S1 A). The immunohistology protocol was followed as previously described (Bao et al., 2017 (link)). The following primary antibodies were used: mouse anti-GFAP (Millipore #MAB360, 1:1,000), mouse anti-NeuN (Millipore #MAB377, 1:1,000), rabbit anti-GABA (Sigma #A2052, 1:500), goat anti-MCM2 (R&D Systems #AF5778, 1:500), chicken anti-Nestin (Aves Labs #NES, 1:1,000), chicken anti-GFP (Aves Labs #GFP-1020, 1:1,000), rabbit anti-RFP (Rockland #600-401-379, 1:1,000), rabbit anti-S100B (Abcam #ab52642, 1:1,000), rat anti-mCherry (Invitrogen #M11217, 1:500), goat anti-DCX (Santa Cruz Biotechnology #sc-8066, 1:100), goat anti-GFP (Rockland #600-101-215, 1:500), and goat anti-SOX2 (Santa Cruz #sc-17320, 1:250). All fluorescent secondary antibodies were obtained from Life Technologies and diluted to 1:500.
All images for quantification were acquired as a z stack, ≤1 μm axial step, then loaded into ImageJ (Fiji) software as a composite image and counted using the Cell Counter plugin. For 3D reconstruction, images were deconvolved using Huygens Professional software (SVI, the Netherlands). 3D reconstruction data were displayed using Surface Renderer in Huygens.
All images for quantification were acquired as a z stack, ≤1 μm axial step, then loaded into ImageJ (Fiji) software as a composite image and counted using the Cell Counter plugin. For 3D reconstruction, images were deconvolved using Huygens Professional software (SVI, the Netherlands). 3D reconstruction data were displayed using Surface Renderer in Huygens.
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