Mouse anti mash1

Manufactured by BD

Sourced in Germany

Mouse anti-Mash1 is a laboratory reagent used to detect and identify the Mash1 protein in biological samples. Mash1 is a transcription factor that plays a role in the development of the nervous system. This antibody can be used in various immunological techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to visualize and quantify the presence of Mash1 in cells and tissues.

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Lab products found in correlation

Goat anti dcx, by Santa Cruz Biotechnology (2 mentions) Rat anti gfap, by Thermo Fisher Scientific (2 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Goat anti iba1, by Abcam (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Cyanine3 (cy3), by Jackson ImmunoResearch (1 mentions) Alexa 488, by Jackson ImmunoResearch (1 mentions) Mouse anti gfap, by Merck Group (1 mentions) Aqua poly mount mounting medium, by Polysciences (1 mentions) Rabbit anti 5 ht, by Immunostar (1 mentions) Mouse anti actub, by Merck Group (1 mentions) Rabbit anti dcx, by Cell Signaling Technology (1 mentions) Rabbit anti sert, by Immunostar (1 mentions) Rabbit anti aciii, by Santa Cruz Biotechnology (1 mentions) Rabbit anti synaptophysin, by Abcam (1 mentions) Rat anti brdu, by Abcam (1 mentions) Chicken anti gfp, by Aves Labs (1 mentions) Mouse anti β catenin, by BD (1 mentions) Goat anti sox2, by Santa Cruz Biotechnology (1 mentions) Rat anti brdu, by Bio-Rad (1 mentions)

8 protocols using mouse anti mash1

Immunohistochemistry Workflow for Neuronal Markers

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Immunohistochemistry was performed as previously published (Sun et al. 2018 (link)). In brief, 3PM imaged mice were perfused and brains were fixed with 4% PFA. Brains were sectioned coronally at 30 μm with a microtome and immunohistochemistry carried out on free-floating sections. Sections were blocked with PBS+ (10% donkey serum, 0.1% Triton-X in PBS) and incubated with primary antibodies overnight at 4 °C, followed with Alexa fluor-conjugated secondary antibodies (Invitrogen). The following primary antibodies were used: rat anti-GFAP (1:500, Invitrogen 130300), goat anti-Dcx (1:100, Santa Cruz sc-8066), goat anti-Iba1 (1:500, Abcam ab5076), rabbit anti-S100ß (Dako Z031129-2), mouse anti-Mash1 (1:100, BD Biosciences 556604). DAPI (Sigma) was used for counterstaining cell nuclei blue before mounting. Zeiss confocal scanning microscopes (Model 710 or 800) were used for image acquisition. About, 1 μm intervals were used for Z-stack scanning.

Sun B., Wang M., Hoerder-Suabedissen A., Xu C., Packer A.M, & Szele F.G. (2022). Intravital imaging of the murine subventricular zone with three photon microscopy. Cerebral Cortex (New York, NY), 32(14), 3057-3067.

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Multimarker Staining of Murine Brain

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Cryosections of brains from 6–7 week old mice were stained with rabbit anti-Ki67 (1:500, Novacastra laboratories, Wetzlar, Germany), goat anti-Dcx (1:100, Santa Cruz biotechnologies), mouse anti-Mash1 (BD Pharmingen). Secondary antibodies coupled with Cy3, Cy5 and Alexa488 (1:300, Jackson ImmunoResearch laboratories, West Grove, USA) were used (see also Supplementary Information Materials and Methods).

Prost G., Braun S., Hertwig F., Winkler M., Jagemann L., Nolbrant S., Leefa I.V., Offen N., Miharada K., Lang S., Artner I, & Nuber U.A. (2016). The putative tumor suppressor gene EphA7 is a novel BMI-1 target. Oncotarget, 7(36), 58203-58217.

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Immunohistochemical Staining Protocol

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Both wholemounts and sections were incubated for 24–48 hours at 4°C with primary antibodies, followed by 24–48 hours at 4°C with the appropriate secondary a ntibodies (Molecular Probes, Invitrogen). The blocking solutions used were PBS/ 0.5% Triton X-100/ 10% Normal Donkey Serum (NDS) for wholemounts, and PBS / 0.1% Triton X-100/ 5% NDS for sections. For BrdU staining, samples were incubated for 40 min at 37°C w ith 2N HCl followed by 10 min at 25°C with 0.1 M boric acid (pH8.5), prior to incubation with antibodies. For mounting, the wholemounts and sections were embedded with Aqua Poly/Mount mounting medium (Polysciences). Primary antibodies: Mouse anti-AcTub (Sigma, 1:1000), Rabbit anti-5HT (Immunostar, 1:500), Rabbit anti-SERT (Immunostar, 1:500), Rabbit anti-synaptophysin (Abcam, 1:500), Goat anti-vGluT3 (Abcam, 1:200), Rabbit anti-ACIII (Santa Cruz, 1:500), Mouse anti-β-catenin (BD Biosciences, 1:500), Chicken anti-GFP (Aves Labs, 1:500), Rabbit anti-DCX (Cell Signaling, 1:200), Mouse anti-GFAP (Millipore, 1:1000), Mouse anti-Mash1 (BD Biosciences, 1:200) and rat anti-BrdU (Abcam, 1:500).

Tong C.K., Chen J., Cebrián-Silla A., Mirzadeh Z., Obernier K., Guinto C.D., Tecott L.H., García-Verdugo J.M., Kriegstein A, & Alvarez-Buylla A. (2014). Axonal Control of the Adult Neural Stem Cell Niche. Cell stem cell, 14(4), 500-511.

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Immunocytochemistry of Frozen Tissue Sections

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Immunocytochemistry of frozen tissue sections was performed essentially as described (Voronova et al., 2017 (link)) and as detailed in the Supplemental Experimental Procedures. The following primary antibodies were used in this study: goat anti-SOX2 (Santa Cruz, 1:250, catalog no. sc-17320), chicken anti-eGFP (Abcam, 1:1,000, catalog no. ab13970), rat anti-GFAP (Invitrogen, 1:300, catalog no. 130300), mouse anti-MASH1 (BD Pharmingen, 1:1,000, catalog no. 556604), rabbit anti-KI67 (Abcam, 1:250, catalog no. ab15580), rabbit anti-DOUBLECORTIN (Cell Signaling Technology, 1:500, catalog no. 4604), rabbit anti-OLIG2 (Millipore, 1:1,000, catalog no. AB9610), goat anti-PDGFRA (R&D Systems, 1:250, catalog no. AF1062), mouse anti-NEUN (Millipore, 1:500, catalog no. MAB377) and rat anti-BrdU (AbD Serotec, 1:300, catalog no. OBT0030). Fluorescently labeled highly cross-absorbed secondary antibodies were purchased from Jackson ImmunoResearch and used at a dilution of 1:1,000. z-Stacked images were collected using a Quorom Spinning Disk confocal microscope system or a Zeiss Axio Imager M2 system with an X-Cite 120LED light source and a C11440 Hamamatsu camera. Images were taken with an optical slice thickness of 0.3 μm and projected z-stacked images are shown.

Storer M.A., Gallagher D., Fatt M.P., Simonetta J.V., Kaplan D.R, & Miller F.D. (2018). Interleukin-6 Regulates Adult Neural Stem Cell Numbers during Normal and Abnormal Post-natal Development. Stem Cell Reports, 10(5), 1464-1480.

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Multi-Label Immunohistochemistry of Brains

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Mice were deeply anesthetized with pentobarbital (50mg/kg) and perfused/fixed with .9% PBS followed by 4% paraformaldehyde. 40 μm coronal sections were cut and standard, multi-label fluorescent immunohistochemistry was performed using the following antibodies: rat anti-BrdU/CldU (1:250, Accurate Chemicals, Westbury, NY, Clone BU 1/75), mouse anti-BrdU/IdU (1:250, BD, Franklin Lakes, NJ, Clone B44), rabbit anti-GFAP (1:2000, Dako, Carpinteria, CA), rabbit anti-TK (from Dr. Michael Sofroniew), sheep anti-EGFR (1:5000, Capralogics, Hardwick, MA), mouse anti-Mash1 (1:250, BD, Franklin Lakes, NJ), rabbit anti-DCX (1:500, Abcam, Cambridge UK), rabbit anti-Iba1 (1:250, Wako, Richmond, VA). Alexafluorophore-conjugated secondary probes (Molecular Probes, Carlsbad, CA) were used. For CldU and IdU staining, antigen-retrieval was performed by first incubating sections in 10mM sodium citrate at >95°C for 15 minutes followed by incubation in 2N HCl at 37°C for 25 minutes.

Thomsen G.M., Le Belle J.E., Harnisch J.A., Mc Donald W., Hovda D.A., Sofroniew M.V., Kornblum H.I, & Harris N.G. (2014). Traumatic brain injury reveals novel cell lineage relationships within the subventricular zone. Stem cell research, 13(1), 48-60.

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Quantifying Protein Expression Using Western Blot

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To determine the efficiency of the shRNAs, we analysed the expression levels of the protein of interest in control and shRNA-treated samples, by SDS–polyacrylamide gel electrophoresis protein separation and immunoblotting. Protein extracts from P19 cells or whole-telencephalon tissue were obtained by lysis with the Promega passive lysis buffer complemented with a cocktail of protease inhibitors (Roche). Western blotting was performed using standard protocols55 (link). The primary antibodies were diluted in the blocking solution at the following concentrations: rabbit anti-Cenpj: 1:400 (Proteintech), rabbit anti-actin, 1:1,000 (Sigma, A2066) and rabbit anti-alpha tubulin, 1:1,000 (Sigma) and mouse anti-Mash1, 1:1,000, BD bioscience. Horseradish peroxidase-conjugated secondary antibodies were used as follows: goat anti-rabbit, 1:200 (Dako, P0449) and goat anti-mouse, 1:1,000 (Sigma, A8924). Uncropped scans of Fig. 1b and Supplementary Fig. 3f blots are shown in Supplementary Fig. 6.

Garcez P.P., Diaz-Alonso J., Crespo-Enriquez I., Castro D., Bell D, & Guillemot F. (2015). Cenpj/CPAP regulates progenitor divisions and neuronal migration in the cerebral cortex downstream of Ascl1. Nature Communications, 6, 6474.

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Profiling Transcription Factor Binding in ESCs

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Trypsin-dissociated ESCs (∼10⁷ cells) were washed in ice-cold PBS and fixed for 10 min at 27°C on a benchtop rotator in neutral buffer containing 1% formalin. Fixed, whole cells were washed, lysed, and sonicated in siliconized microfuge tubes in an ice-chilled Diagenode Bioruptor for a total of 35 min, using a 15:15 sec on/off cycle. Chromatin immunoprecipitation was performed with an overnight incubation at 4°C with 5 µg of mouse anti-MASH1 (BD Pharmingen 556604) for ASCL1 ChIP or mouse anti-FLAG (Sigma-Aldrich F1804) antibody for ASCL2 and MYOD1 ChIP. The FLAG fusion moiety in ESCs with inducible ASCL1 was not recognized by FLAG antibodies, so antibodies specific to ASCL1 were used (Nishiyama et al. 2009 (link)). Bound DNA fragments were isolated after 4–6 h incubation at 4°C with 25 µg Protein G Dynabeads (Life Technologies 10003D). Purified samples were tested for enrichment of previously identified regions by RT-qPCR. Duplicate ChIP purified samples were pooled prior to library preparation to generate a sufficient template for library generation. Samples were prepared as ChIP libraries using NEB Next library preparation kit and Illumina multiplexing primers. Samples were sequenced using an Illumina HiSeq 2500 line.

Casey B.H., Kollipara R.K., Pozo K, & Johnson J.E. (2018). Intrinsic DNA binding properties demonstrated for lineage-specifying basic helix-loop-helix transcription factors. Genome Research, 28(4), 484-496.

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Pituitary Cell Immunostaining

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Immunostaining was performed with the following antibodies as previously described: rabbit anti-POMC (ACTH) (1:4000; Chemicon), rabbit anti-GH (1:4000; Chemicon), rabbit anti-PRL (1:1000; Chemicon), mouse anti-TSH (1:50; Dako), mouse anti-FSH (1:50; Dako), mouse anti-LH (1:50; Dako), mouse anti-Mash1 (1:500; BD Pharmingen), goat anti-NeuroD (1:200; Santa Cruz), rabbit anti-SOX2 (1:2000; Abcam), rabbit anti-GFP

Ando M., Goto M., Hojo M., Kita A., Kitagawa M., Ohtsuka T., Kageyama R, & Miyamoto S. (2018). The proneural bHLH genes Mash1, Math3 and NeuroD are required for pituitary development. Journal of molecular endocrinology, 61(3).

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