Sc 8066

Manufactured by Merck Group

The SC-8066 is a laboratory equipment product manufactured by Merck Group. It is a high-precision instrument designed for specific applications within the scientific research and testing fields. The core function of the SC-8066 is to perform accurate measurements and analyses as required by the user's research or testing protocols. Further details on the intended use or specific applications of this product are not available.

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Lab products found in correlation

Mab377, by Merck Group (2 mentions) Mab377, by Agilent Technologies (1 mentions) Rat anti ki67, by Thermo Fisher Scientific (1 mentions) Z0311, by Agilent Technologies (1 mentions) Nel701001kt, by PerkinElmer (1 mentions) Cy3 conjugated donkey anti goat, by Jackson ImmunoResearch (1 mentions) Cy5 conjugated donkey anti mouse, by Jackson ImmunoResearch (1 mentions) Vt1000, by Leica (1 mentions) Sliding microtome, by Reichert Technologies (1 mentions) Alexa 488 donkey anti goat, by Thermo Fisher Scientific (1 mentions) Micro cover glasses, by Avantor (1 mentions) Sc 8066, by Santa Cruz Biotechnology (1 mentions) Alexa conjugated secondary antibodies, by Jackson ImmunoResearch (1 mentions) Af3075, by R&D Systems (1 mentions) Ab9610, by R&D Systems (1 mentions) A11122, by Thermo Fisher Scientific (1 mentions) Apotome system, by Zeiss (1 mentions) Gfp 1020, by Santa Cruz Biotechnology (1 mentions) Hoechst 33342, by Merck Group (1 mentions) Anti neurofilament, by Santa Cruz Biotechnology (1 mentions)

6 protocols using sc 8066

Immunofluorescence Protocol for Brain Sections

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Immunofluorescence was performed as previously described [17 (link)]. Briefly, brains were sectioned at a thickness of 30 µm and left 1 h in blocking solution (10% normal donkey serum, 0.3% Triton-X1000, 0.04% NaN₃ in PBS). Following the blocking step, the sections were incubated with primary antibodies (diluted in blocking solution) overnight at 4 °C. The primary antibodies used were rabbit anti-Ascl1 (Cosmo Bio Co., CAC-SK-T01-003, concentration 1:250), goat anti-Dcx (Santa Cruz, SC-8066, concentration 1:500), mouse anti-NeuN (Millipore, MAB377, concentration 1:500), rabbit anti-S100β (DAKO, Z0311, concentration 1:200, antibody was directly conjugated with fluorophore Alexa-647) and rat anti-Ki67 (eBioscience, 14-5698, concentration 1:1000). Subsequently, the sections were washed three times for 5 min in PBS before incubation with secondary antibodies (1:500 in blocking solution) against the species for the specific primary antibodies and conjugated with the fluorophores Alexa-488, Cy3 and Alexa-647. For Ascl1 detection we used a Tyramide Signal Amplification (TSA) protocol (PerkinElmer, NEL701001KT).

Santopolo G., Magnusson J.P., Lindvall O., Kokaia Z, & Frisén J. (2020). Blocking Notch-Signaling Increases Neurogenesis in the Striatum after Stroke. Cells, 9(7), 1732.

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Immunohistochemical Visualization of Adult Neurogenesis

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Mice were deeply anesthetized with ketamine/xylazine (150/15 mg/kg) and transcardially perfused with cold 0.1 M phosphate buffered saline (PBS), followed by 25mL of cold 4% paraformaldehyde in PBS. Brains were postfixed overnight in 4% paraformaldehyde at 4°C, then cryoprotected in 30% sucrose in PBS, and stored at 4°C. Coronal sections (35 µm) were cut through the entire hippocampus on a cryostat and stored in PBS with 0.1% NaN₃.
For doublecortin (DCX) immunohistochemistry, sections were blocked in 10% normal donkey serum in 0.1M phosphate-buffered saline (PBS) with 0.25% Triton X-100 for 2h at room temperature. After overnight incubation with primary antibodies (goat anti-DCX, 1:500, Santa Cruz #SC 8066; mouse anti-NeuN 1:500, Millipore MAB377), sections were washed in PBS and incubated with secondary antibodies (Cy3-conjugated donkey anti-goat and Cy5-conjugated donkey anti-Mouse, 1:500, Jackson ImmunoResearch) for 2h at room temperature. Sections were mounted on slides, dried, rinsed in water, and cover slipped using aqueous mounting medium.

Swan A.A., Clutton J.E., Chary P.K., Cook S.G., Liu G.G, & Drew M.R. (2014). Characterization of the role of adult neurogenesis in touch-screen discrimination learning. Hippocampus, 24(12), 1581-1591.

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Avian Neuronal Immunolabeling Protocol

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Immediately after capture (still on the field) and under deep isoflurane anesthesia, birds were perfused transcardially with phosphate buffered saline 0.1M followed by aldehyde fixatives (4% paraformaldehyde, 0.1 M phosphate buffer, pH 7.2–7.4). Brains were dissected, stored in phosphate buffered saline 0.1M and cut by Vibratome (Leica VT1000S) or freezing microtome (Sliding Microtome, Reichert Jung) in the coronal plane. Six anatomical series of sections (6 parallel series), cut at 60 or 80μm in the freezing microtome or vibratome respectively, were immunolabeled for NeuN or DCX or stained by Nissl. Free-floating sections were immunolabeled with anti-doublecortin antibody (Santa Cruz SC-8066) or NeuN antibody (MiliporeSigma, MAB377), and mounted on glass slides coated with an aqueous solution of gelatin (10%) and chromium potassium sulfate (0.5%). Sections were air-dried at room temperature, dehydrated, cleared in alcohol/xylene series and covered with 50% entellan (Entellan® Novo 107961, Merck Milipore) diluted in xylene and cover slipped.

de Morais Magalhães N.G., Guerreiro Diniz C., Guerreiro Diniz D., Pereira Henrique E., Corrêa Pereira P.D., Matos Moraes I.A., Damasceno de Melo M.A., Sherry D.F, & Wanderley Picanço Diniz C. (2017). Hippocampal neurogenesis and volume in migrating and wintering semipalmated sandpipers (Calidris pusilla). PLoS ONE, 12(6), e0179134.

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Peptide Competition Assay for DCX Antibodies

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To determine the specificity of the goat anti-DCX antibody (Santa Cruz, sc-8066) and the guinea-pig anti-DCX antibody (Chemicon, AB5910) a peptide competition assay was performed. The DCX peptide was no longer available at Chemicon. As such, both antibodies were submitted to a competition assay with the DCX peptide provided by Santa Cruz Biotechnology (sc-8066). A 1:5 solution of anti-DCX antibody and anti-DCX peptide (sc-8066P, Santa Cruz) was incubated at room temperature for 1 h. The preparation was then diluted in 0.3% Triton and each section was covered with 50 μl and incubated at room temperature for 3 h. Following incubation, slides were washed successively three times for 5 min in PBS (10 mM). Each section was then incubated for 30 min at 37°C with 50 μl of the secondary antibody Alexa488 donkey anti-Goat (1:1,000, Invitrogen) or Alexa488 anti-guinea pig (1:500, Jackson Immuno Research) diluted in 0.3% Triton in 10X PBS. After incubation, slides were washed successively three times for 5 min in 10X PBS. Sections were imbedded in custom-made anti-fade solution (p-Phenylenediamine and glycerol in PBS solution) and cover-slipped with micro cover glasses (VWR Scientific).

Boulanger J.J, & Messier C. (2017). Doublecortin in Oligodendrocyte Precursor Cells in the Adult Mouse Brain. Frontiers in Neuroscience, 11, 143.

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Immunofluorescent Labeling of Brain Sections

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Mice were transcardially perfused with 4% paraformaldehyde. The brains were removed, post-fixed overnight, and cut into 50 μm coronal and sagittal sections using a vibratome (Leica Microsysteme, Rueil Malmaison, France). Immunofluorescent labeling was performed on sections or on cultured cells fixed with paraformaldehyde 4%. The following antibodies were used: anti-GFP (rabbit, 1/500; Life Technologies, A11122. Chicken, 1/500; Aves Labs GFP-1020), anti-DCX (goat, 1/100; Santa Cruz Sc-8066), anti-MBP (mouse IgG1, 1/500; Chemicon MAB384), anti-GFAP (mouse IgG 1/1,000; Sigma G3893), anti-OLIG2 (rabbit, 1/500; Chemicon AB 9610), anti-SOX9 (goat, 1/200; R&D AF3075), anti-PDGFRα (rat, 1/250; Chemicon CBL1366), anti-CC1 (mouse, 1/500; Calbiochem OP80), anti-CASPR/PARANODIN (rabbit, 1/500; L51, gift from Dr Goutebroze), anti-TCF4 (mouse IgG2a, 1/500; Millipore 05–511), anti-PSA-NCAM (mouse, ½; supernatant produced in our laboratory), anti-NEUROFILAMENT-160 (mouse, 1/500; Sigma N5264), and anti-βIV-SPECTRIN (mouse clone 393/2, 1/500; NeuroMab AB_2315816). The sections and cells were incubated with appropriate Alexa-conjugated secondary antibodies (1/500; Jackson ImmunoResearch Laboratories) then counterstained with Hoechst 33,342 (1/1,000; Sigma). Images were captured with a Zeiss apotome system (20× and 60× objectives) and a Zeiss 510 confocal (60× objective).

El Waly B., Cayre M, & Durbec P. (2018). Promoting Myelin Repair through In Vivo Neuroblast Reprogramming. Stem Cell Reports, 10(5), 1492-1504.

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Immunostaining and Confocal Imaging Protocol

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Slice preparation, immunostaining, and image acquisition and analysis were as previously described (Platel et al., 2009 (link)). Primary antibodies included: anti-VGAT1 (mouse, 1:500, Synaptic Systems), anti-neurofilament (NF, 1:500, MAB5166) anti-DCX (goat or rabbit, 1:100, Santa Cruz, SC8066 and SC28939), and anti-GLAST (guinea pig, 1:500, Chemicon). Each staining was replicated at least in 4–5 slices from three different mice. The appropriate secondary antibodies were Alexa fluor series (1:1000, Invitrogen, USA) or Cyanine series (1:500, Jackson Labs). Z-section images (spaced by 1–2 μm over 10–20 μm) were acquired on a laser-scanning confocal microscope (Olympus FluoView 1000) with a 20× dry objective (N.A. 0.75) or a 60× oil objective (N.A. 1.42). Images were analyzed and reconstructed using Imaris 4.0 (Bitplane AG, Switzerland) and Photoshop CS3 (Adobe, USA).

Young S.Z., Lafourcade C.A., Platel J.C., Lin T.V, & Bordey A. (2014). GABAergic striatal neurons project dendrites and axons into the postnatal subventricular zone leading to calcium activity. Frontiers in Cellular Neuroscience, 8, 10.

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