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Cells were washed with cold PBS and total proteins were extracted with RIPA buffer containing Halt inhibitors (Thermofisher, MA)
Standard injection protocol: Dose: 1 to 2 mg daily Duration: 10 to 20 day cycles Frequency: 1 to 3 cycles per year, spaced 4 to 6 months apart Timing: Evening administration to align with natural melatonin production timing Administration: Subcutaneous or intramuscular Maintenance protocol (sublingual): Daily sublingual sprays between injection cycles Lower bioavailability than injection but convenient for ongoing support The cyclic nature of Epitalon protocols reflects how the peptide works
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