Immunostaining of Muscle Cells

First, monoclonal and polyclonal antibodies, including MyoG (sc-12732, 1:150, Santa Cruz), MEF2C (#5030 s, 1:200, CST), and MyHC (sc-20641, 1:150, Santa Cruz), were added to each well in every group and then incubated for 12 h at 4 °C. The incubated cells were washed with PBS 3 times for 15 min and subsequently treated with the appropriate fluorescent dye-labeled secondary antibodies (Jackson Lab, 1:500, USA) at 25 °C for 2 h. The nuclei were stained with DAPI (Molecular Probes). The images for each group were photographed under a Nikon 80i fluorescence microscope^{18 (link)}.

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Yue J., Xu W., Xiang L., Chen S.J., Li X.Y., Yang Q., Zhang R.N., Bao X., Wang Y., Mbadhi M., Liu Y., Yao L.Y., Chen L., Zhao X.Y., Hu C.Q., Zhang J.X., Zheng H.T., Wu Y., Chen S.Y., Li S., Lv J., Shi L.L, & Tang J.M. (2023). Continuous exposure to isoprenaline reduced myotube size by delaying myoblast differentiation and fusion through the NFAT-MEF2C signaling pathway. Scientific Reports, 13, 436.

Publication 2023

fluorescent dye-labeled secondary antibodies DAPI Nikon 80i fluorescence microscope

Antibodies Cells Dapi Fluorescence Fluorescent dye Molecular probes Nuclei

Corresponding Organization : Taihe Hospital

Other organizations : Zunyi Medical University, University of Missouri

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Variable analysis

independent variables

Monoclonal and polyclonal antibodies, including MyoG (sc-12732, 1:150, Santa Cruz), MEF2C (#5030 s, 1:200, CST), and MyHC (sc-20641, 1:150, Santa Cruz)

dependent variables

Unclear from the information provided

control variables

Incubation time (12 h at 4 °C)
Washing with PBS (3 times for 15 min)
Incubation with secondary antibodies (25 °C for 2 h)
Staining with DAPI (Molecular Probes)
Imaging with Nikon 80i fluorescence microscope

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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