Myosin heavy chain

Manufactured by Thermo Fisher Scientific

Myosin Heavy Chain is a structural protein found in muscle cells. It plays a crucial role in muscle contraction by interacting with actin filaments. The Myosin Heavy Chain is responsible for the generation of the force required for muscle movement.

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Lab products found in correlation

Myogenin, by Thermo Fisher Scientific (2 mentions) Cfx96 real time system thermocycler, by Bio-Rad (2 mentions) Rneasy mini kit, by Qiagen (2 mentions) Iscript cdna synthesis kit, by Bio-Rad (2 mentions) Taqman gene expression master mix, by Thermo Fisher Scientific (1 mentions) Taqman fast universal pcr master mix without amperase ung, by Thermo Fisher Scientific (1 mentions) Myod1, by Thermo Fisher Scientific (1 mentions)

2 protocols using myosin heavy chain

Quantitative PCR Analysis of Myogenic Markers

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To further support immunostaining data, quantitative PCR was performed on sub-confluent and differentiated cells from P2 and P6. Briefly, RNA was harvested with the RNEasy Mini Kit (Qiagen #74104, Hilden, Germany) and cDNA was prepared using 200 ng of RNA for each reaction with the iScript cDNA synthesis kit (Bio-Rad # 1708890, Hercules, CA, USA) according to the manufacturers’ instructions. Next, qPCR was performed using the TaqMan Gene Expression Master Mix (ThermoFisher #4369016), with 18 S as a housekeeping control gene. Primers used were: 18 S (ThermoFisher #Hs03003631), Pax3 (ThermoFisher #Bt04303789), MyoD (ThermoFisher #Bt04282788), Myogenin (ThermoFisher #Bt03258929), and Myosin Heavy Chain (ThermoFisher #Bt03273061). Reactions were performed using 3.5 uL of the prepared cDNA, according to the manufacturer’s instructions. Reactions were run on the Bio-Rad CFX96 Real Time System thermocycler, and results were analyzed as 2^−Δct normalized to 18 S expression.

Stout A.J., Mirliani A.B., Rittenberg M.L., Shub M., White E.C., Yuen JS J.r, & Kaplan D.L. (2022). Simple and effective serum-free medium for sustained expansion of bovine satellite cells for cell cultured meat. Communications Biology, 5, 466.

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Quantifying BSC Gene Expression

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To assess relative gene expression between BSCs cultured in various media types, qPCR was performed on proliferative (70% confluency) and differentiated cells from P3 following standard protocols. Briefly, RNA was harvested using an RNEasy Mini kit (Qiagen #74104, Hilden, Germany) and cDNA was prepared via the iScript cDNA synthesis kit (Bio-Rad #1708890, Hercules, CA, USA) using 1 μg of RNA for each reaction. Next, qPCR was performed using 2 μL of cDNA and 1x TaqMan Fast Universal PCR Master Mix without AmpErase UNG (ThermoFisher #4352042). Primers used in this study were: 18S (ThermoFisher #Hs03003631), Pax3 (ThermoFisher #Bt04303789), MyoD1 (ThermoFisher #Bt03244740), Myogenin (ThermoFisher #Bt03258929), and Myosin Heavy Chain (ThermoFisher #Bt03273061). Reactions were performed according to the manufacturer’s instructions on a Bio-Rad CFX96 Real Time System thermocycler, and results were analyzed as 2^−ΔΔct normalized to expression of the 18S housekeeping gene and analyzed relative to proliferative BSC-GM expression for each gene.

Stout A.J., Rittenberg M.L., Shub M., Saad M.K., Mirliani A.B., Dolgin J, & Kaplan D.L. (2023). A Beefy-R culture medium: replacing albumin with rapeseed protein isolates. Biomaterials, 296, 122092.

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