Muscle Enzyme Activity Measurement

The maximal activities of CS, β‐HAD, and COX were measured as described elsewhere [10 (link), 17 (link)]. Frozen and crushed gastrocnemius muscles were homogenized in 100 times (vol/wt) of 100 mm potassium phosphate buffer using a μT‐01 bead crusher (TAITEC, Saitama, Japan). Maximal enzyme activity was measured spectrophotometrically. The total protein concentration in the homogenates was determined using a BCA protein assay (TaKaRa BIO Inc., Shiga, Japan), with activities normalized to total protein concentration.

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Takahashi K., Kitaoka Y., Matsunaga Y, & Hatta H. (2021). Lactate administration does not affect denervation‐induced loss of mitochondrial content and muscle mass in mice. FEBS Open Bio, 11(10), 2836-2844.

Publication 2021

BCA protein assay

Assay Buffer Enzyme activity Frozen Gastrocnemius muscles Potassium phosphate Protein

Corresponding Organization : The University of Tokyo

Other organizations : Kanagawa University

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Variable analysis

dependent variables

Maximal activities of CS, β-HAD, and COX

control variables

Frozen and crushed gastrocnemius muscles
Homogenization in 100 times (vol/wt) of 100 mM potassium phosphate buffer using a μT-01 bead crusher
Measurement of maximal enzyme activity spectrophotometrically
Determination of total protein concentration in the homogenates using a BCA protein assay
Normalization of activities to total protein concentration

positive controls

None specified

negative controls

None specified

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