Medium was replaced with Seahorse XF Base Medium (Agilent) supplemented with 15 mM glucose (Gibco, A2494001), 1 mM sodium pyruvate (Gibco, 11360070), and 4 mM L-glutamine (Gibco, 25030081), pH 7.4, unless indicated otherwise and incubated in a non-CO2 incubator at 37 C for less than one hour
This supplement is made with hypoallergenic, vegan ingredients and comes in a serving size of one capsule with sixty servings per container
Sundaresan, S
It contains two unnatural amino acids alpha-aminoisobutyric acid (Aib) at position 1 and D-2-naphthylalanine (D-2-Nal) at position 3 along with a D-phenylalanine at position 4
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Add 1,000 L (1 mL) of reconstitution solvent to the 1 mg vial This yields a 1 mg/mL (1,000 g/mL) stock solution To achieve 100 g/mL working stock: Dilute 100 L of 1 mg/mL stock into 900 L of PBS or cell culture medium yields 100 g/mL in 1 mL To achieve 10 ng/mL in a 5 mL experiment: Take 0.5 L of 100 g/mL working stock + 4,999.5 L of medium Alternatively: dilute to an intermediate concentration to reduce pipetting error Use a dilution series approach (stock working stock final experimental concentration) to minimize pipetting inaccuracy at nanogram-level concentrations
