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Skeletal muscle myosin heavy chain (MyHC) fiber type composition is a critical determinant of overall muscle function and health. Various approaches interrogate fiber type at the single cell, but the two most commonly utilized are single muscle fiber sodium dodecyl sulfate polyacrylamide gel electrophoresis (smfSDS-PAGE) and fluorescent immunohistochemistry (IHC). Although smfSDS-PAGE is generally considered the "gold standard", IHC is more commonly used due to its time-effectiveness and relative ease. Unfortunately, there is lingering inconsistency on how best to accurately and quickly determine fiber type via IHC, and an overall misunderstanding regarding pure fiber type proportions, specifically the abundance of fibers exclusively expressing highly-glycolytic MyHC IIX in humans. We therefore: 1) present information and data showing the low abundance of pure MyHC IIX muscle fibers in healthy human skeletal muscle and, 2) leverage this information to provide straightforward protocols that are informed by human biology and employ inexpensive, easily attainable antibodies for the accurate determination of fiber type.
This article was published in the following journal.
Name: Journal of applied physiology (Bethesda, Md. : 1985)
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The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results.
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