Fastprep 24 5g instrument

The frozen yeast pellet is resuspended in 1 mL of cold double-distilled water (ddH₂O) and 250 μL of glass microbeads. Yeast cells were broken by a high-speed benchtop homogenizer (MP Biomedicals™ FastPrep-24™ 5G Instrument, Fisherbrand, Waltham, MA, USA) at 6.5 m/s with 5 cycles and a 60 s pause in-between. Then, the samples were centrifuged at 10,000 rpm for 10 min, and the supernatants were used to perform the antioxidant assays. The reported protocols are a variation of the methods compiled by Kesraoui et al. [30 (link)] (namely: total phenolics [31 ], reducing power [32 (link)], ferrous ion chelating activity [33 (link)], and β-carotene-linoleic acid assay [34 (link)]). During the current protocols, only the key variations and the concise process are presented in Section 2.6, Section 2.7, Section 2.8 and Section 2.9. ddH₂O was used as the negative control of all the assays. In order to be certain that the variation in antioxidant assays was due to real biological changes, the results were normalized by the value at t = 0 h.

One millilitre (1 mL) of Invitrogen™ TRI Reagent™ Solution per 50 mg to 100 mg of gonadal white adipose tissue (gWAT), inguinal white adipose tissue (iWAT), intercapsular brown adipose tissue (iBAT) and pericardial adipose tissue (PAT) was transferred to a MP Biomedicals^TM Lysing Matrix D tube. Lysing tubes were placed in the MP Biomedicals™ FastPrep-24™ 5G Instrument, with the QuickPrep Adaptor (Fisher Scientific) and homogenised at a speed of 6.0 m/s, for 40 s. Isolation of RNA was performed as recommended by the manufactures of Invitrogen™ TRI Reagent™ Solution (Fisher Scientific).