Mab377

Manufactured by Fujifilm

Sourced in United States, Germany

The MAB377 is a lab equipment product manufactured by Fujifilm. It is designed to perform key laboratory functions. The core functionality of the MAB377 is to assist in the analysis and processing of samples within a controlled laboratory environment.

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Lab products found in correlation

Ab4674, by Abcam (2 mentions) Eclipse ti2, by Nikon (2 mentions) Ab13970, by Abcam (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Alexa fluor 568, by Thermo Fisher Scientific (2 mentions) Papain, by Thermo Fisher Scientific (1 mentions) Optiprep, by Merck Group (1 mentions) Neuronal nuclei (neun), by Merck Group (1 mentions) Hibernate a, by Thermo Fisher Scientific (1 mentions) Alexa fluor 633, by Thermo Fisher Scientific (1 mentions) Anti iba1, by Fujifilm (1 mentions) Ab40390, by Abcam (1 mentions) Ne1023, by Merck Group (1 mentions) Tcs sp8, by Leica (1 mentions) Ab290, by Abcam (1 mentions) Mouse anti gfap cy3, by Merck Group (1 mentions) Rabbit anti iba1, by Fujifilm (1 mentions) Fv1000 confocal laser scanning microscope, by Olympus (1 mentions) Mouse anti neun, by Merck Group (1 mentions) Abc reagent, by Vector Laboratories (1 mentions)

9 protocols using mab377

Isolation and Characterization of Neuronal and Microglial Populations from Aging Mouse Brains

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Adult neurons and microglia were isolated from the mice brain as previously described^{11 (link)}. In brief, the cortex of WT (1, 3, 5, or 9-months-old), APP/PS1 (1, 3, 5, or 9-months-old), APP/PS1 treated with N-AS (9-months-old) or APP/PS1 treated with aspirin (9-months-old) mice were minced in Hibernate A (Gibco)/B27 (Invitrogen) medium and dissociated using papain (Worthington) solution. After tissue trituration, cells were separated by Optiprep (Sigma-Aldrich) density gradient centrifugation. The purity of the fractionated neurons and microglia were acutely confirmed by neuronal (NeuN, mouse, 1:200, Millipore, MAB377) and microglial (Iba1, rabbit, 1:1000, Wako, 019-19941) markers and the isolated neurons and microglia were acutely analyzed for the acetylation assay and lipid mediators metabololipidomics.

Lee J.Y., Han S.H., Park M.H., Song I.S., Choi M.K., Yu E., Park C.M., Kim H.J., Kim S.H., Schuchman E.H., Jin H.K, & Bae J.S. (2020). N-AS-triggered SPMs are direct regulators of microglia in a model of Alzheimer’s disease. Nature Communications, 11, 2358.

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Immunofluorescent Labeling of Brain Tissue

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The removed brain tissue was cryoprotected using optimal cutting temperature compound (OCT, Tissue-Tek, Torrance CA), frozen, and sectioned. Tissue sections were hydrated in PBS and blocked with 10% normal goat serum. Following 45 min in 0.5% Triton X-100 in PBS, the sections were incubated overnight with primary antibodies for mouse monoclonal NeuN (Millipore, MAB377, 1:500), rabbit polyclonal anti-Iba1 (Wako Chemicals, 01919741, 1:500), rabbit monoclonal caspase-3 (Cell Signaling, 9661S, 1:500), chicken GFAP (Sigma, AB5541, 1:500) and DAPI. Sections were then rinsed three times for five minutes each with PBS and incubated for two hours with AlexaFluor488 (goat anti rabbit, 1:500), AlexaFluor568 (goat anti mouse 1:500), and AlexaFluor633 (goat anti chicken, 1:500), from ThermoFisher Scientific (Waltham, Massachusetts, USA). Images were acquired with an Olympus FluoView 1000 confocal fluorescent microscope (Olympus, Inc., Tokyo, Japan) at the Center for Biologic Imaging at the University of Pittsburgh.

Robbins E.M., Castagnola E, & Cui X.T. (2022). Accurate and stable chronic in vivo voltammetry enabled by a replaceable subcutaneous reference electrode. iScience, 25(8), 104845.

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Quantifying White Matter Damage in Mouse Brain

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Coronal tissue sections were incubated with one of the primary antibodies: anti‐MBP antibody (1:300, ab40390, Abcam, UK), SMI‐32 (1:200, NE1023, Millipore, USA), Neuron (1:100, Millipore, MAB377, USA), or Iba 1 (1:1000, Wako, 019–19,741, USA) in a blocking solution at 4°C overnight. Subsequently, sections were exposed to secondary antibodies, specifically DAM‐Alexa488 and DAR‐Alexa555, and incubated at room temperature for 2 h. Fluorescent images were captured using a confocal laser scanning microscope (TCS SP8, Leica, Germany). The quantitative analysis involved assessing the immunostaining intensity of both MBP and SMI32 antibodies, as well as enumerating target immunopositive cells using ImageJ software.
²⁶ (link) A total of three randomly chosen microscopic fields from each of the three sequential sections were examined per mouse brain. White matter damage was quantitatively determined by calculating the average intensity ratio of SMI32 to MBP.

Ye L., Tang X., Zhong J., Li W., Xu T., Xiang C., Gu J., Feng H., Luo Q, & Wang G. (2024). Unraveling the complex pathophysiology of white matter hemorrhage in intracerebral stroke: A single‐cell RNA sequencing approach. CNS Neuroscience & Therapeutics, 30(3), e14652.

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Multimodal Mapping of Brain Injury

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Triplicate 10 μm thick brain sections 0, 0.5, and 1 mm away from the needle tract were counterstained with Hoechst using standard protocols. Additional sections 0.5 mm from the needle tract were stained with mouse anti-NeuN (1 : 800, Millipore MAB377), rabbit anti-Iba1 (1 : 500, Wako 019-19741), and chicken anti-GFAP (1 : 500, Abcam ab4674) using standard protocols. Fluorescent images were obtained on a Nikon Eclipse Ti2 (Nikon Instruments Inc.) at 10x magnification for distribution studies in whole brain slices. Distribution images were thresholded in ImageJ to obtain particle distribution, which was then quantified by area and represented as a schematic image with overlapping replicates. Cell localization images were obtained at 40x magnification in the striatum.

Waggoner L.E., Miyasaki K.F, & Kwon E.J. (2023). Analysis of PEG-lipid anchor length on lipid nanoparticle pharmacokinetics and activity in a mouse model of traumatic brain injury. Biomaterials Science, 11(12), 4238-4253.

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Fluorescent Immunohistochemistry for Viral Vector Expression

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Fluorescent immunohistochemistry was used to visualise viral vector gene expression in which cells the gene expression was occurring. As determined by DAB immunohistochemistry, sections were selected that had highest vector expression in the series. The sections were blocked and permeabilised with 10% goat serum in PBST (0.2%). Sections were incubated with primary antibodies; rabbit-anti-dYFP 1:20,000 (Abcam #ab290), mouse-anti-dYFP 1:200 (Abcam #ab291), mouse-anti-GFAP-Cy3 1:1000 (Sigma #C9205), mouse-anti-NeuN 1:250 (Merck #MAB377), rabbit-anti-Iba1 1:250 (Wako #019–19741), rabbit-anti-olig2 diluted 1:500 in PBST + 4% goat serum overnight at room temperature and then incubated with complementary secondary antibodies; all AlexaFluor diluted 1:750 in PBST + 4% goat serum for 4 h at room temperature. Sections were mounted onto positively charged glass slides and coverslipped with prolong gold. After 2 days of curing, the slides were imaged on an Olympus FV1000 confocal laser scanning microscope at ×60 magnification. Sections from every animal used in the study were stained.

Griffin J.M., Fackelmeier B., Fong D.M., Mouravlev A., Young D, & O’Carroll S.J. (2019). Astrocyte-selective AAV gene therapy through the endogenous GFAP promoter results in robust transduction in the rat spinal cord following injury. Gene Therapy, 26(5), 198-210.

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Neuroinflammation Profiling in Brain Injury

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Triplicate 10 μm thick brain sections 0, 0.5, and 1 mm away from the needle tract were counterstained with Hoechst using standard protocols. Additional sections 0.5 mm from the needle tract were stained with mouse anti-NeuN (1:800, Millipore MAB377), rabbit anti-Iba1 (1:500, Wako 019-19741), and chicken anti-GFAP (1:500, Abcam ab4674) using standard protocols. Fluorescent images were obtained on a Nikon Eclipse Ti2 (Nikon Instruments Inc.) at 10x magnification for distribution studies in whole brain slices. Distribution images were thresholded in ImageJ to obtain particle distribution, which was then quantified by area and represented as a schematic image with overlapping replicates. Cell localization images were obtained at 40x magnification in the striatum.

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Immunohistochemical Analysis of Mouse Brain

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Three-month-old male mice were anesthetized and transcardially perfused with 0.9% saline containing heparin (10 mg/L) followed by fixative (4% paraformaldehyde, 15% picric acid, 0.1% glutaraldehyde in 0.1 M PB). Brains were collected and post-fixed overnight before coronal sections were taken at every 50 μm. Sections were washed and then treated with 1% H₂O₂ for 15 min to remove endogenous peroxidase activity. After washing and blocking with 2% normal horse serum, sections were incubated with primary antibodies [anti-mouse GFAP, 1:1000 for 2 h at room temperature (RT), Sigma, G3893; anti-rabbit c-fos, 1:2000 for overnight at RT, Millipore, ABE457; anti-chicken GFP, 1/2000 for overnight at RT, Abcam, ab13970; anti-mouse NeuN, 1/1000 for overnight at RT, Millipore, MAB377; anti-rabbit Iba-1, 1/2000 for overnight at RT, Wako, 019-19741]. The following day, sections were extensively washed and incubated in biotinylated anti-rabbit secondary antibody, ABC reagent, and diaminobenzidine (DAB) substrate (Vector Laboratories). Crystal violet staining was performed to detect cell nuclei. Immunofluorescence was performed with a combination of Alexa Fluor 488–or Alexa Fluor 594–labeled anti-rabbit, anti-chicken, or anti-mouse secondary antibody (1:500 for 1 h at RT, Invitrogen). Representative images were selected from at least 3 times repeated experiments.

Kim1 J.G., Suyama S., Koch M., Jin S., Argente-Arizon P., Argente J., Liu Z.W., Zimmer M.R., Jeong J.K., Szigeti-Buck K., Gao Y., Garcia-Caceres C., Yi C.X., Salmaso N., Vaccarino F.M., Chowen J., Diano S., Dietrich M.O., Tschöp M.H, & Horvath T.L. (2014). Leptin signaling in GFAP-expressing adult glia cells regulates hypothalamic neuronal circuits and feeding. Nature neuroscience, 17(7), 908-910.

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Multiplexed Immunofluorescence Labeling of Neuronal Subtypes

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To assess brain cellular subtype markers (i.e., neurons, microglia, astrocytes, and oligodendrocytes) co-expressed with EGFP, striatal and hippocampal monkey sections were incubated simultaneously with two antibodies: a chicken polyclonal antibody against GFP (Abcam, ab13970, 1:1000) and, respectively, a mouse monoclonal antibody against NeuN (Millipore, Darmstadt, Germany, MAB377, 1:1000) or a rabbit monoclonal against human Iba1 (FUJIFILM Wako Pure Chemical Corporation, 019-19741, 1:1000) or a mix of mouse monoclonal antibodies against GFAP and S-100 (Sigma-Aldrich (clone GA5), 1:2000/Abcam, ab7852, 1:1000) or a mouse monoclonal antibody against CNPase (Abcam, ab237961, 1:1000) for one night at room temperature. Incubation with secondary antibodies was done sequentially with a goat anti-chicken AlexaFuor 488 (Abcam, A11039, 1:400) and a goat anti-mouse AlexaFluor 568 (Invitrogen, A11031, 1:400) or a goat anti-rabbit AlexaFluor 568 (Invitrogen, A11036, 1:400) 1h30 at room temperature. Sections were mounted on non-gelatinized slides and coverslipped using fluorescent mounting media without DAPI (Vector Labs). Images were acquired using a Zeiss SP5 confocal microscope.

Arotcarena M.L., Dovero S., Biendon N., Dutheil N., Planche V., Bezard E, & Dehay B. (2021). Pilot Study Assessing the Impact of Intrathecal Administration of Variants AAV-PHP.B and AAV-PHP.eB on Brain Transduction in Adult Rhesus Macaques. Frontiers in Bioengineering and Biotechnology, 9, 762209.

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Immunohistochemical Characterization of Neural Cells

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Specimens were fixed in 4% paraformaldehyde/1X PBS at 4 °C for 24 h (TL/TLE cases), and up to 72 h in the case of germinal matrix, rinsed in 1X PBS, and vibratome sectioned (30 µm). Sections were incubated for 1 h in blocking solution (1X PBS/0.5% Triton X-100/10% normal donkey serum); then for 24 h at 4 °C in primary antibody (1X PBS/0.25% Triton X-100/1% normal donkey serum); and then for 4 h at room temperature in either donkey anti-mouse, donkey anti-rabbit, donkey anti-rat, or donkey anti-goat fluorochrome-conjugated secondary antibodies (Jackson Laboratories, 1:250 dilution). Formalin-fixed paraffin embedded (FFPE) tissues underwent 1 h deparaffinization, rehydration in decreasing gradient of ethanol, and antigen retrieval for 20 min prior to blocking. Primary antibodies dilutions were as follows: 1:50 mouse anti-EGFR (Invitrogen 280005); 1:500 rat anti-GFAP (Life Technologies, 13–0300); 1:250 rabbit anti-OLIG2 (Millipore, AB9610); 1:250 goat anti-hOLIG2 (R&D Systems, AF2418SP); 1:250 rabbit anti-Ki67 (Abcam, Ab15580); 1:250 mouse anti-Ki67 (BD Biosciences, 556003); 1:100 rabbit anti-PAX6 (Novus Biologicals, NBP1-89100); 1:100 mouse anti- NEUN (Millipore, MAB377); 1:250 rabbit anti-AIF1 (IBA1) (Wako, 019–19741). Nuclei were counterstained with DAPI (1:1000). Images were obtained using a LSM 780 upright confocal microscope (Zeiss).

Pai B., Tome-Garcia J., Cheng W.S., Nudelman G., Beaumont K.G., Ghatan S., Panov F., Caballero E., Sarpong K., Marcuse L., Yoo J., Jiang Y., Schaefer A., Akbarian S., Sebra R., Pinto D., Zaslavsky E, & Tsankova N.M. (2022). High-resolution transcriptomics informs glial pathology in human temporal lobe epilepsy. Acta Neuropathologica Communications, 10, 149.

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