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After a final wash step, a tetramethylbenzidine (TMB) peroxidase substrate solution (KPL Inc.) was added to each well and absorbance of the developed color was measured by a colorimetric plate reader (Molecular Device)

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

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Visceral Fat: Specific targeting of visceral adipocytes which have high GH receptor density