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Membranes were stripped for 20 min at room temperature using a Blot Restore Membrane Rejuvenation kit (Sigma-Aldrich, 2520-M), then washed twice with TBS-T, re-blocked with BSA 5% for 30 min and incubated with mouse-anti--actin (Sigma-Aldrich, A1978) 1:3,000 in BSA 2.5% for 4 h at room temperature and then incubated with a sheep-anti-mouse (Amersham, NA931) secondary antibody at 1:5,000 in BSA 2.5% for 2 h

Absolute Contraindications Active malignancy Pregnancy and lactation Severe hypoglycemia disorders Known hypersensitivity to IGF-1 Active proliferative retinopathy Relative Contraindications Diabetes mellitus Renal impairment Cardiac impairment History of hyperplasia Family history of certain cancers Severe hepatic impairment Dosing & Administration Starting Dose 2050 mcg/day subcutaneously Begin at the lower end for sensitive patients Assess tolerance for 12 weeks before titration Therapeutic Range 50120 mcg/day for most indications Maximum dose: 200 mcg/day Individualize dosing based on response and tolerance Cycling Protocol 46 weeks on therapy 4 weeks off therapy Helps prevent receptor desensitization Supports long-term efficacy Administration Guidelines Hypoglycemia Prevention: Always administer IGF-1 LR3 with or shortly after carbohydrate-containing meals

Storage Instructions: Store at room temperature or refrigerated