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Identification of Skeletal Muscle Satellite Cells by ...

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6100681/
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Apr 19, 2018 ... Immunofluorescence is an effective method that helps to identify ... The above protocol was based on a method of Pax7/MF20 staining on ... US National Library of Medicine Search databaseSearch termSearch COVID-19 is an emerging, rapidly evolving situation. Journal ListJ Vis Exp(134); 2018PMC6100681 Published online 2018 Apr 19. doi: 10.3791/57212 Identification of Skeletal Muscle Satellite Cells by Immunofluorescence with Pax7 and Laminin Antibodies Xuesong Feng,# 1 Faiza Naz,# 2 Aster H. Juan, 1 Stefania Dell'Orso, 2 and Vittorio Sartorelli 1 Immunofluorescence is an effective method that helps to identify different cell types on tissue sections. In order to study the desired cell population, antibodies for specific cell markers are applied on tissue sections. In adult skeletal muscle, satellite cells (SCs) are stem cells that contribute to muscle repair and regeneration. Therefore, it is important to visualize and trace the satellite cell population under different physiological conditions. In resting skeletal muscle, SCs reside between the basal lamina and myofiber plasma membrane. A commonly used marker for identifying SCs on the myofibers or in cell culture is the paired box protein Pax7. In this article, an optimized Pax7 immunofluorescence protocol on skeletal muscle sections is presented that minimizes non-specific staining and background. Another antibody that recognizes a protein (laminin) of the basal lamina was also added to help identify SCs. Similar protocols can also be used to perform double or triple labeling with Pax7 and antibodies for additional proteins of interest. Keywords: Developmental Biology, Issue 134, Skeletal muscle, satellite cells, Pax7, immunofluorescence, stem cell, basal lamina



MYH1E Antibody (MF 20) - DSHB

https://dshb.biology.uiowa.edu/MF-20
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mouse myosin, sarcomere (MHC) MYH1E antibody validated for ELISA,FACS, FFPE,Immunofluorescence,Immunohistochemistry,Immunoprecipitation,Western  ...---



Preparation and Culture of Myogenic Precursor Cells/Primary ...

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5408649/
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Feb 16, 2017 ... Here, we show a simple experimental protocol for robust, ... Seed 50,000 cells/ well in a 12-well plate for MF 20, senescence and viability assays. ... in block 2); for MF 20 staining, use MYH1E (MF 20) primary antibody (DSHB, ... US National Library of Medicine Search databaseSearch termSearch COVID-19 is an emerging, rapidly evolving situation. Journal ListJ Vis Exp(120); 2017PMC5408649 Published online 2017 Feb 16. doi: 10.3791/55047 Preparation and Culture of Myogenic Precursor Cells/Primary Myoblasts from Skeletal Muscle of Adult and Aged Humans Ana Soriano-Arroquia, 1 Peter D. Clegg, 1 Andrew P. Molloy, 1 , 2 and Katarzyna Goljanek-Whysall 1 This article has been cited by other articles in PMC. Skeletal muscle homeostasis depends on muscle growth (hypertrophy), atrophy and regeneration. During ageing and in several diseases, muscle wasting occurs. Loss of muscle mass and function is associated with muscle fiber type atrophy, fiber type switching, defective muscle regeneration associated with dysfunction of satellite cells, muscle stem cells, and other pathophysiological processes. These changes are associated with changes in intracellular as well as local and systemic niches. In addition to most commonly used rodent models of muscle ageing, there is a need to study muscle homeostasis and wasting using human models, which due to ethical implications, consist predominantly of in vitro cultures. Despite the wide use of human Myogenic Progenitor Cells (MPCs) and primary myoblasts in myogenesis, there is limited data on using human primary myoblast and myotube cultures to study molecular mechanisms regulating different aspects of age-associated muscle wasting, aiding in the validation of mechanisms of ageing proposed in rodent muscle. The use of human MPCs, primary myoblasts and myotubes isolated from adult and aged people, provides a physiologically relevant model of molecular mechanisms of processes associated with muscle growth, atrophy and regeneration. Here we describe in detail a robust, inexpensive, reproducible and efficient protocol for the isolation and maintenance of human MPCs and their progeny — myoblasts and myotubes from human muscle samples using enzymatic digestion. Furthermore, we have determined the passage number at which primary myoblasts from adult and aged people undergo senescence in an in vitro culture. Finally, we show the ability to transfect these myoblasts and the ability to characterize their proliferative and differentiation capacity and propose their suitability for performing functional studies of molecular mechanisms of myogenesis and muscle wasting in vitro.



Myosin 4 Monoclonal Antibody (MF20), Alexa Fluor 488, eBioscience

https://www.thermofisher.com/antibody/product/...4...MF20.../53-6503-82
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Invitrogen Anti-Myosin 4 Monoclonal (MF20), eBioscience™, Catalog # 53-6503- 82. Tested in Immunofluorescence (IF), Immunocytochemistry (ICC) and ... Myosin 4 Monoclonal Antibody (MF20), Alexa Fluor 488, eBioscience™ View all (5) Myosin 4 antibodies Check your price & availability Usually ships in 1 business day Eye (London, England) 2016 - Published figure using Myosin 4 monoclonal antibody (Product # 53-6503-82) By clicking this image or link above, you will be redirected to the Benchsci website. Disclaimer Immunohistochemistry (Frozen) (IHC (F)) Immunohistochemistry (Paraffin) (IHC (P)) Bovine, Dog, Chicken, Chimpanzee, Cat, Guinea pig, Human, Mouse, Non-human primate, Rabbit, Rat Mouse IgG2b kappa Isotype Control (eBMG2b), Alexa Fluor 488, eBioscience™ Alexa Fluor® 488 See Additional Formats 4° C, store in dark, DO NOT FREEZE! Description: This MF20 monoclonal antibody recognizes the heavy chain of myosin II, specificially the light meromyosin portion, in cardiac and skeletal muscle of vertebrates. Myosin II is composed of two heavy chains and four light chains. The 220-kDa myosin heavy chain exists as four different isoforms due to alternative splicing. Myosins interact with actin and hydrolyze ATP to function in muscle contraction, cytokinesis, and phagocytosis. The MF20 has been shown to react to myosin from... View more Muscle contraction. Weiss A., J. Mol. Biol. 290:61-75(1999). Denis N. J., Proteomics 7:868-874(2007). For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization. Have you cited this antibody in a publication? Let us know so we can reference it in this datasheet.



Myosin Heavy Chain Antibody (MF20) [Unconjugated] (MAB4470 ...

https://www.novusbio.com/.../myosin-heavy-chain-antibody-mf20_mab4470
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Feb 25, 2015 ... Mouse Monoclonal Anti-Myosin Heavy Chain Antibody (MF20) [Unconjugated] ... MYLK3 Antibody [NBP2-62682] - Staining of human heart. Support: +31 800 0225607 | info.emea@bio-techne.com Home » Myosin Heavy Chain » Myosin Heavy Chain Antibodies » Myosin Heavy Chain Antibody (MF20) [Unconjugated] Myosin Heavy Chain Antibody (MF20) [Unconjugated] (49 Publications) (1 Review) See All Related Submit a Review Myosin Heavy Chain was detected in immersion fixed paraffin-embedded sections of human skeletal muscle using Mouse Anti-Myosin Heavy Chain Monoclonal Antibody (Catalog # MAB4470) at 5 µg/mL for 1 hour at room ...read more Reactivity All-MultiSpecies Glossary View Available Conjugates View Available Formulations 100% Guarantee on every product Datasheet Reviews & Publications Protocols & FAQs Support & Research Myosin Heavy Chain Antibody (MF20) [Unconjugated] Summary Chicken pectoralis-derived Myosin Detects Myosin Heavy Chain in human, mouse, rat and other mammalian, avian, and amphibian species. Test in a species/application not listed above to receive a full credit towards a future purchase. Learn about the Innovator's Reward Immunohistochemistry 5-25 ug/mL MAB4470 in the following applications: MAB4470 in the following applications: Packaging, Storage & Formulations Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 6 months, -20 to -70 °C under sterile conditions after reconstitution. Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.




A Method for the Direct Identification of Differentiating Muscle Cells ...

https://journals.plos.org/plosone/article?id=10.1371/journal.pone...
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Dec 9, 2011 ... MitoTracker staining provides a robust and simple detection strategy for ... with MF-20 (primary antibody for MyHC) or F5D (for MyoG) (DSHB, ... Best Practices in Research Reporting MANUSCRIPT REVIEW AND PUBLICATION Editorial and Peer Review Process Discover a faster, simpler path to publishing in a high-quality journal. PLOS ONE promises fair, rigorous peer review, broad scope, and wide readership – a perfect fit for your research every time. LEARN MORE SUBMIT NOW ABOUT A Method for the Direct Identification of Differentiating Muscle Cells by a Fluorescent Mitochondrial Dye Tetsuaki Miyake , John C. McDermott, Anthony O. Gramolini Identification of differentiating muscle cells generally requires fixation, antibodies directed against muscle specific proteins, and lengthy staining processes or, alternatively, transfection of muscle specific reporter genes driving GFP expression. In this study, we examined the possibility of using the robust mitochondrial network seen in maturing muscle cells as a marker of cellular differentiation. The mitochondrial fluorescent tracking dye, MitoTracker, which is a cell-permeable, low toxicity, fluorescent dye, allowed us to distinguish and track living differentiating muscle cells visually by epi-fluorescence microscopy. MitoTracker staining provides a robust and simple detection strategy for living differentiating cells in culture without the need for fixation or biochemical processing. Editor: Antonio Paolo Beltrami, University of Udine, Italy Received: August 2, 2011; Accepted: November 11, 2011; Published: December 9, 2011 Funding: This study was supported by grants from the Heart and Stroke Foundation (HSF) of Ontario (#T-6281; to AOG), Canadian Institutes of Health Research (CIHR) (to AOG; MOP-84267), and Canadian Foundation for Innovation to AOG. AOG is a Canada Research Chair and a New Investigator of the HSF of Canada. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.






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