Cryosections of urethral tissue samples were prepared as described previously[15 (link)]. Immunofluorescence was performed as described previously [16 (link)] using phalloidin and anti-myosin heavy chain antibody (Abcam Inc., Cambridge, MA, USA). To identify intramyocellular lipid (IMCL) within urethral striated muscle, we employed the staining procedure reported previously [13 (link)]. After washing with PBS, the slides were incubated with LipidTOX neutral lipid stain (1:1000, Invitrogen) at room temperature for 30 minutes, followed by staining with phalloidin (Invitrogen). For image analysis, five randomly selected fields per tissue per animal for each treatment group were photographed and recorded using a Retiga Q Image digital still camera and ACT-1 software (Nikon Instruments Inc., Melville, NY, USA).
Anti myosin heavy chain antibody
Manufactured by Abcam
Sourced in United States
The Anti-myosin heavy chain antibody is a laboratory reagent used to detect and measure the presence of myosin heavy chain, a structural protein found in muscle cells. This antibody can be used in various experimental techniques, such as immunohistochemistry, Western blotting, and ELISA, to study the expression and distribution of myosin heavy chain in biological samples.
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Lab products found in correlation
Act 1, by Nikon (1 mentions)
Lipidtox neutral lipid stain, by Thermo Fisher Scientific (1 mentions)
Retiga q image digital still camera, by Nikon (1 mentions)
Alexa fluor 647, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions)
Meta confocal microscope with zen software, by Zeiss (1 mentions)
2 protocols using anti myosin heavy chain antibody
1
Urethral Muscle Lipid Staining Protocol
2
Myogenic Differentiation Assay
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To confirm myogenic differentiation, freshly fused DEC (n = 4 fusions) and parent myoblasts (snj MBwt and MBmdx) were cultured on German glass coverslips in serum-free Myogenic Differentiation Medium (Promocell, Heidelberg, Germany) supplemented with 10 μg/ml insulin to induce myogenic differentiation. After seven-day culture, cells were fixed with ice-cold acetone and unspecific antibody binding was blocked with 10% normal goat serum. Rabbit polyclonal anti-myosin heavy chain antibody (1:200, Abcam, Cambridge, MA, USA) was used as primary and goat anti-rabbit Alexa Fluor 647 (1:500, Molecular Probes, OR, USA) conjugated antibody was used as secondary antibody. Nuclei were counterstained with DAPI Vector Laboratories, CA, USA. A Zeiss Meta confocal microscope with ZEN software (Carl Zeiss, Oberkochen, Germany) was used for fluorescence signal detection and analysis.
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