The pellet was subsequently washed with 10 ml of the G-solution, which comprised of 500 ml HBSS (BE10-508F, Life Technologies) with 10% BSA (126615, Sigma Aldrich) and 1% Pen-Strep (15140122, Life Technologies), and re-suspended in 5.5 ml of gradient solution (15% Optiprep (5 ml 10% RPMI, Life Technologies) + 3 ml of 40% Optiprep, which was diluted from 60 % Optiprep with G-solution (D1556, Sigma Aldrich)) per sample, and placed on top of 2.5 ml of the gradient solution
Cycling GHK-Cu reduces total peptide used, limits theoretical copper accumulation and often produces stronger response when you resume
Nausea is the most common initial side effect, typically mild and improving by day 7 as your body adjusts
When food intake drops significantly, so does your intake of essential micronutrients
(PubMed) Michaud DS, Spiegelman D, Clinton SK, Rimm EB, Willett WC, Giovannucci EL