Mouse anti neun

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom, Japan, Canada, Israel, China

Mouse anti-NeuN is a primary antibody that recognizes the neuronal nuclear antigen (NeuN), a protein expressed in the nuclei and cytoplasm of most neuronal cell types. It is commonly used as a marker for identifying and quantifying neurons in various tissues and applications.

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Lab products found in correlation

Rabbit anti iba1, by Fujifilm (44 mentions) Mouse anti gfap, by Merck Group (43 mentions) Rabbit anti gfap, by Agilent Technologies (25 mentions) Rabbit anti cleaved caspase 3, by Cell Signaling Technology (18 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (16 mentions) Alexa fluor 488, by Thermo Fisher Scientific (13 mentions) Rabbit anti gfp, by Thermo Fisher Scientific (13 mentions) Chicken anti gfp, by Abcam (13 mentions) Rabbit anti gfap, by Merck Group (13 mentions) Mouse anti map2, by Merck Group (12 mentions) Triton x 100, by Merck Group (11 mentions) Hoechst 33342, by Thermo Fisher Scientific (10 mentions) Goat anti chat, by Merck Group (9 mentions) Rabbit anti ki67, by Abcam (9 mentions) Rabbit anti gfap, by Abcam (8 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (8 mentions) Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (8 mentions) Goat anti dcx, by Santa Cruz Biotechnology (8 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (8 mentions) Rat anti brdu, by Bio-Rad (8 mentions)

339 protocols using mouse anti neun

Cellular Expression of TrkB and Signaling

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1) To assess the cellular source of TrkB, double immunofluorescence labeling was performed by simultaneous incubation of sections with rabbit anti-TrkB (1:100; Cell Signaling Danvers, MA, USA) overnight at 4 °C with mouse anti-NeuN (a neuronal marker; 1:100; Millipore, Billerica, MA, USA), rat anti-GFAP (an astrocyte marker; 1:200; Invitrogen, Camarillo, CA, USA), and mouse anti-F4/80 (a microglia/macrophage marker; 1:100; Serotec, Düsseldorf, Germany).
2) To assess protein expression of pTrkB, pAkt Ser473 or BDNF, double immunofluorescence labeling was performed by simultaneous incubation of sections with rabbit anti-pTrkB (1:100; Cell Signaling), rabbit anti-pAkt Ser473 (1:100; Cell Signaling) or rabbit anti-BDNF (1:100; Santa Cruz, CA, USA) overnight at 4 °C with mouse anti-NeuN (1:100; Millipore) or rat anti-GFAP (1:200; Invitrogen).
Sections were washed, followed by incubation with Alexa Fluor 488- or Alexa Fluor 594-conjugated secondary antibodies (1:500; Molecular Probes, Eugene, OR, USA) for 2 h.

Wu C.H., Chen C.C., Hung T.H., Chuang Y.C., Chao M., Shyue S.K, & Chen S.F. (2019). Activation of TrkB/Akt signaling by a TrkB receptor agonist improves long-term histological and functional outcomes in experimental intracerebral hemorrhage. Journal of Biomedical Science, 26, 53.

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Immunohistochemical Analysis of TLR2 and TLR4 in Brain

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The paraffin-embedded brain was serially cut into 10 μm-thick sections in coronal plane. After the standard histological procedures, sections were incubated overnight at 4°C in the mixture of 1:100 rabbit anti-TLR2 (Boster Biotechnology Company, Wuhan, China)/1:100 mouse anti-NeuN (Millipore Corporation, Billerica, USA), 1:100 rabbit anti-TLR2/1:1,000 mouse anti-GFAP (Millipore Corporation, Billerica, USA), 1:100 rabbit anti-TLR2 (Boster Biotechnology Company, Wuhan, China), 1:100 rabbit anti-TLR4 (Boster Biotechnology Company, Wuhan, China)/1:100 mouse anti-NeuN (Millipore Corporation, Billerica, USA), 1:100 mouse anti-TLR4 (Abcam, Cambridge, USA)/1:1,000 rabbit anti-GFAP (Millipore Corporation, Billerica, USA), 1:100 mouse anti-TLR4 (Abcam, Cambridge, USA). Then the sections were incubated respectively with 1:800 Alexa Fluor 568 donkey anti-rabbit (Invitrogen Life Technologies, NY, USA), 1:300 Alexa Fluor 555 donkey anti-mouse (Beyotime) for one hour at room temperature, followed incubated with 1:800 Alexa Fluor 488 donkey anti-mouse (Invitrogen Life Technologies, NY, USA), 1:800 Alexa Fluor 488 donkey anti-rabbit (Invitrogen Life Technologies, NY, USA), 1:100 Lectin-FITC conjugate (Sigma-Aldrich, St. Louis, USA) for onehour at room temperature. Finally, these sections were observed under laser confocal microscope (Leica, Germany).

Wang Y., Ge P., Yang L., Wu C., Zha H., Luo T, & Zhu Y. (2014). Protection of ischemic post conditioning against transient focal ischemia-induced brain damage is associated with inhibition of neuroinflammation via modulation of TLR2 and TLR4 pathways. Journal of Neuroinflammation, 11, 15.

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Immunofluorescent Quantification of Neural Markers

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Three sections (1.21 mm rostral and 0.23 mm and 1.31 mm caudal to bregma) from each mouse were washed with 1X PBS, blocked with 10% Bovine Serum Albumin (BSA) for at least 1 hour, and then incubated overnight at 4 °C with mouse anti-NeuN (Millipore) and 1:100 rabbit anti-cleaved caspase-3 (Cell Signaling), or mouse anti-NeuN (Millipore) and rabbit anti-LC3B (Sigma). The sections were then incubated with 1:200 AlexaFluor 546 goat anti-mouse and AlexaFluor 488 goat anti-rabbit (Invitrogen), or 1:200 AlexaFluor 488 rabbit anti-mouse and AlexaFluor 546 donkey anti-rabbit (Invitrogen) for one hour at room temperature. Finally, sections were mounted with DAPI mounting medium with Vector shield and visualized using a Nikon fluorescence microscope (Eclipse Ts2). For quantitative analysis, both NeuN and cleaved caspase-3 or LC3B positive cells were counted in one parietal cortex area and two temporal cortex areas surrounding the infarct core per section and expressed as percentage change to the control group.

Li C., Li J., Xu G, & Sun H. (2020). Influence of Chronic Ethanol Consumption on Apoptosis and Autophagy Following Transient Focal Cerebral Ischemia in Male Mice. Scientific Reports, 10, 6164.

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Immunohistochemical Analysis of Spinal Cord Injury

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Frozen coronal sections (7 μm) were collected near the collagenase injection site and washed three times in PBS at room temperature. Antigen retrieval was performed by placing slides in a pressure cooker containing citrate buffer (pH 6.0) for 4 min. Tissues were permeabilized by 1% triton-X100 in PBS for 30 min, washed in PBS, and blocked in 1% BSA in PBS for 1 hr. Slides were incubated overnight at 4°C in 1% BSA containing the following primary antibodies: mouse anti-3-NT (1:500, Sigma), rabbit anti-iNOS (1:300, Cell Signaling), rabbit anti-GFAP (1:1000, Abcam), mouse anti-Iba1 (1:1000, Wako), rabbit anti-CD31 (1:300, EMD Millipore), and mouse anti-NeuN (1:1000, Sigma). The slides were washed in PBS, and then incubated for 1 hr at room temperature in 1% BSA containing Alexa Fluor®594-conjugated goat anti-rabbit and DyLight^™488-conjugated goat anti-mouse (1:500, Jackson ImmunoResearch) secondary antibodies. After a final PBS wash, slides were mounted with SlowFade® Diamond Antifade Mountant with DAPI (ThermoFisher) and a coverslip. Images were captured using fluorescent microscopy. Mean fluorescence intensity and cell count per field were quantified by a person who was blinded to the experimental groups, using NIH image J software.

Bahader G.A., Nash K.M., Almarghalani D.A., Alhadidi Q., McInerney M.F, & Shah Z.A. (2021). Type-I Diabetes Aggravates Post-Hemorrhagic Stroke Cognitive Impairment by augmenting Oxidative Stress and Neuroinflammation in Mice. Neurochemistry international, 149, 105151.

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Quantifying Mitochondrial Morphology in Neurons

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Sections fixed as described above were blocked with 0.5% Triton X-100 + 5% normal donkey serum in PBS for 1 h at RT, then incubated with rabbit anti-TOM20 (1:50, Santa Cruz Biotechnology, #sc-11415), mouse anti-NeuN (1:1000, Sigma-Aldrich, #MAB377), or rabbit anti-GFAP (1:500, Dako, #Z0334) O/N at 4°C. After rinsing, sections were incubated with donkey anti-rabbit or anti-mouse Alexa 555 (1:500, Thermo Fisher Scientific, #A31572 and #A31570) for 2 h at RT.
To mark nuclei, fixed slices were incubated with the nuclear marker TO-PRO 3 (1:10000, Sigma-Aldrich) for 10 min at RT and rinsed in PBS.
Slices were mounted and coverslipped with Mowiol, then imaged using a Leica TCS SP5 II confocal system equipped with a HCX PL Fluotar 20×/0.50 or a HCX PL APO 100×/1.40 objective. The Argon laser line (488 nm) was used to excite fluorescent proteins (FPs) of the Cameleon sensor. Confocal microscopy imaging was performed at 1024 × 1024 pixels per image, with a 100 Hz acquisition rate, average of two frames.
The analysis of the area occupied by mitochondria expressing the probe was performed on 3 slices from 3 animals using ImageJ software (US National Institutes of Health). For each image, a threshold was imposed and the fluorescent area was determined.

Redolfi N., Greotti E., Zanetti G., Hochepied T., Fasolato C., Pendin D, & Pozzan T. (2021). A New Transgenic Mouse Line for Imaging Mitochondrial Calcium Signals. Function, 2(3), zqab012.

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Immunohistochemical Analysis of Amyloid Plaques

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Animals were deeply anesthetized, killed, and perfused intracardially with normal saline and fixed with 4% paraformaldehyde (PFA). Brains were placed in 15% sucrose in PBS for 6-12 h, then 30% sucrose in PBS until equilibrated. Brain coronal sections (10 μm) were cut with a cryostat (Leica CM1900), sections were used for immunohistochemistry. Slices were washed with PBS and fixed with 4% paraformaldehyde, permeabilized with 0.05% Triton X-100, then pre-incubated in 1% BSA solution for blocking. Slices were incubated overnight at 4 °C with the primary antibody solution (rabbit anti-amyloid-β antibody, Abcam; 1:500), mouse anti-NeuN (1:100, Sigma), rabbit anti-glial fibrillary acidic protein (GFAP) (1:250, Sigma) on a shaker. After washing with PBS, the sections were incubated with the secondary antibody (goat anti-rabbit Alexa Fluor 488 antibody, Abcam; 1:1000; goat anti-mouse Alexa Fluor 488 antibody, Abcam; 1:1000) for 2 h at room temperature. Sections were mounted in the vectashield mounting medium with DAPI (Vector laboratories). For the detection of amyloid plaques, brain tissue sections were stained in 0.1% thioflavin-S (Sigma) and rinsed with 70% ethanol. The brain tissue sections were also stained with hematoxylin and eosin (H&E) to assess the histological damage.

Deng Z., Wang J., Xiao Y., Li F., Niu L., Liu X., Meng L, & Zheng H. (2021). Ultrasound-mediated augmented exosome release from astrocytes alleviates amyloid-β-induced neurotoxicity. Theranostics, 11(9), 4351-4362.

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Quantitative Analysis of TET2 Protein in DRG

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As reported before [32 (link)], the mice were anesthetized with 1% pentobarbital sodium and then perfused with normal saline followed by a solution of 4% paraformaldehyde (PFA). The DRGs were post-fixed in 4% PFA for 4 h and then dehydrated sequentially in 20% and 30% sugar solutions. Each dehydrated DRG sample was excised into 10-µm-thick serial sections, which were then mounted on glass slides. After antigen retrieval and blocking with 5% fetal bovine serum, the DRG sections were co-incubated overnight at 4 °C with rabbit anti-TET2 antibody and DAPI with one of the following antibodies: mouse anti-NeuN (1:500, Sigma), mouse anti-NF200 (1:500, Sigma), or mouse anti-CGRP (1:500, Abcam). Subsequently, after washing the slides three times with phosphate buffered saline (PBS) (for 5 min each time), the sections were incubated with a mixture of Alexa Fluor 488 goat anti-rabbit IgG (H + L) and Alexa Fluor 568 goat anti-mouse IgG (H + L) (1:200, Invitrogen, Life technologies™, USA) 1 h in dark condition. Images were captured under a fluorescence microscope. We acquired at least 9 locations in the DRG per mouse: we quantified 3 images per section and 3 sections per mouse. We quantified at least 3 mice per experiment blindly.

Chen W., Wang X., Sun Q., Zhang Y., Liu J., Hu T., Wu W., Wei C., Liu M., Ding Y., Liu D., Chong Y., Wang P., Zhu H., Cui W., Zhang J., Li Q, & Yang F. (2022). The upregulation of NLRP3 inflammasome in dorsal root ganglion by ten-eleven translocation methylcytosine dioxygenase 2 (TET2) contributed to diabetic neuropathic pain in mice. Journal of Neuroinflammation, 19, 302.

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Quantifying Neuronal Expression in Hippocampal Tissue

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Neuronal membranes of fixed hippocampi were permeated using 1 mL of a permeabilization (wash) buffer (200 mL PBS [Invitrogen], 200 μL Triton X-100 [Sigma], 0.010 mg Bovine Serum [Sigma]) for 45 minutes at room temperature. Hippocampi were exposed to primary monoclonal antibody mouse anti-NeuN (Sigma) at a dilution factor of 1:200 for 24 hours at 4°C. Hippocampi were then washed twice with 1x PBS and exposed to goat anti-mouse secondary antibody fluorescein isothiocyanate (FITC; Sigma) at a dilution factor of 1:200 for 24 hours at 4°C. Hippocampi were washed twice with 1x PBS and imaged immediately via SPOT software 4.0.2 (advanced version) for Windows (W. Nuhsbaum Inc.; McHenry, IL, USA) through a 5x objective with a Leica DMIRB microscope (W. Nuhsbaum Inc.; McHenry, IL, USA). Images were captured with a SPOT 7.2 color mosaic camera (W. Nuhsbaum). Densitometry using Image J software (National Institutes of Health, Bethesda, MD) was conducted to measure the intensity of FITC fluorescence. FITC fluorescence was detected with a band-pass filter at 495 nm (520 nm emission). Background measurements were recorded for each reading and subtracted from the measurements obtained from each primary cell layer of the hippocampal formation (the granule cell layer of the dentate gyrus and the pyramidal cell layers of the CA3 and CA1 regions).

Reynolds A.R., Williams L.A., Saunders M.A, & Prendergast M.A. (2015). Group 1 mGlu-Family Proteins Promote Neuroadaptation to Ethanol and Withdrawal-Associated Hippocampal Damage. Drug and alcohol dependence, 156, 213-220.

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Immunofluorescence Staining of Neural Markers

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The following primary antibodies were used: anti-BrdU monoclonal antibodies (1:400, Oxford Biotechnology, UK), goat anti-DCX (1:400; Santa Cruz Biotechnology, CA, USA), rabbit anti-BDNF (1:200; Abcam, Cambridge, MA, USA), goat anti-mouse IGF-1 (1:200; BD Bioscience, San Jose, CA, USA), rat anti-mouse anti-CD3 (eBioscience, Santa Clara, CA, USA), rabbit anti-Iba-1 (1:1000; Wako Chemical, Japan), mouse anti-GFAP (1:10,000; Sigma-Aldrich, St. Louis, MO, USA), mouse anti-IL-10 (1:200; BD Bioscience, San Jose, CA, USA), chicken anti-Arginase-1 (1:500; Merck Millipore, Germany), rat anti-CD11b (1:1000; BD Bioscience, San Jose, CA, USA), and mouse anti-NeuN (1:1000; Sigma-Aldrich, St. Louis, MO, USA). The following secondary antibodies were used: Alexa Fluor 594 donkey anti-rat, Alexa Fluor 488 donkey anti-goat, Alexa Fluor 555 goat anti-rabbit, Alexa Fluor 488 goat anti-rabbit, Alexa Fluor FITC goat anti-chicken, and Alexa Fluor 488 goat anti-mouse antibodies (1:400; Molecular Probes, Eugene, OR, USA). The detailed immunofluorescence protocols were previously described [23 (link)].

Qi F., Zuo Z., Yang J., Hu S., Yang Y., Yuan Q., Zou J., Guo K, & Yao Z. (2017). Combined effect of BCG vaccination and enriched environment promote neurogenesis and spatial cognition via a shift in meningeal macrophage M2 polarization. Journal of Neuroinflammation, 14, 32.

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Multimodal Neuronal Tissue Imaging

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Mice were transcardially perfused with 0.9% saline followed by 4% paraformaldehyde using a peristaltic pump (Gilson) and fixed overnight at 4 °C. The tissue was then dehydrated using 30% sucrose until it sank to the bottom of tube. Tissue sections were taken on a Leica CM1950-Cryostat (Leica) at a thickness of 40 μm for brain, 10 μm for liver. Sections were rinsed three times in 0.1M phosphate buffer (PB) and incubated with the primary antibodies: rabbit anti-mCherry (1:3 000, GeneTex), chicken anti-GFP (1:1000,Invitrogen),mouse anti-NeuN (1:3000,Sigma), which were diluted in diluent with 5% NGS overnight at 4 °C. The following day, sections were washed three times in PB and then incubated with the secondary antibodies: 561-AffiniPure Goat Anti-Rabbit IgG (1:500,Jackson Immunoresearch), 488-AffiniPure Goat Anti-Chicken IgG (1:500, Invitrogen) and Cy5-AffiniPure Goat Anti-Mouse IgG (1:500, Jackson Immunoresearch) for 2 h at room temperature on an orbital shaker. Finally, the sections were counterstained with DAPI for 20 min and mounted with SlowFade Diamond Antifade Mountant (Life) on glass slides.

Yao X., Wang X., Hu X., Liu Z., Liu J., Zhou H., Shen X., Wei Y., Huang Z., Ying W., Wang Y., Nie Y.H., Zhang C.C., Li S., Cheng L., Wang Q., Wu Y., Huang P., Sun Q., Shi L, & Yang H. (2017). Homology-mediated end joining-based targeted integration using CRISPR/Cas9. Cell Research, 27(6), 801-814.

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