We obtained blood from mouse tail veins and measured overnight-fasted blood glucose biweekly with a Contour glucometer (Contour; Bayer). In addition, glucose tolerance tests were performed before sacrifice to determine insulin resistance (Supplemental Figure 3 ). Plasma cholesterol and triglyceride levels were determined with enzymatic assay kits (Wako Chemicals) using blood obtained by cardiac puncture. Liver toxicity was assessed by measuring AST and ALT activity in plasma using enzymatic activity assay kits (Abcam). We used dual-energy X-ray absorptiometry (DEXA) analysis to assess fat mass and lean body mass before each sacrifice.
Enzymatic activity assay kits
Manufactured by Abcam
Enzymatic activity assay kits are laboratory tools designed to measure the activity of specific enzymes. They provide a standardized and quantitative method to assess the functional capacity of enzymes in various biological samples. These kits include all the necessary reagents and protocols to perform the assay, enabling researchers to determine enzyme kinetics and activity levels.
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2 protocols using enzymatic activity assay kits
1
Metabolic Profiling in Mouse Models
2
Enzymatic Activities of Cathepsins B and D
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Enzymatic activities of cathepsin B and cathepsin D were determined in snap-frozen DD cords and nodules from the same 3 patients used in the WB, using enzymatic activity assay kits for cathepsin B (cat# ab65300; Abcam) and cathepsin D (cat# ab65302; Abcam). All steps of the procedure were performed according to the manufacturer’s protocol. Fluorescence was measured in a Nunc F96 MicroWell black polystyrene plate (cat# 136101, Thermo Fisher Scientific) using the Varioskan Flash plate reader (cat# MIB5250030, Thermo Fisher Scientific). Tonsil and denatured tonsil tissue lysates were used as appropriate positive and negative controls, respectively.
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