Pur a lyzer mini dialysis kit

0.05 mg ATTO 488 N-hydroxysuccinimidyl (NHS) ester (ATTO-Tech, Singen, Germany) dissolved in DMSO was added to 0.5 mg of the coactivator-derived peptide (JPT, Berlin, Germany) with a final DMSO concentration of 1%. The reaction was carried out for one hour at room temperature and was quenched with 100 mM Tris, pH 7.5^{25 (link),44 (link)}. Free label was removed by dialysis against 20 mM Tris, 20 mM NaCl, 5 mM MgCl₂, pH 7.5 using a Pur-A-Lyzer Mini Dialysis Kit (Sigma, St. Louis, USA) with a 1 kDa molecular weight cut-off.

The in vitro drug release study was carried out using the Pur-A-Lyzer Mini Dialysis Kit and the molecular weight cut off (MWCO) of the dialysis membrane was 3.5 Kda (Sigma-Aldrich, St. Louis, MO, USA). The study was performed in a shaking incubator at 100 rpm and 37 °C. The carvedilol loaded transfersome suspension containing 0.2 mg carvedilol or the same amount of carvedilol in acetone (the acetone volume was the same as the suspension) was added into the dialysis tubes. The dialysis tubes were immersed in 200 mL of PBS (pH 7.4) as the release medium. At predetermined time intervals (0.5, 1, 2, 3, 4, 6, 8, 24 h), 1 mL samples were withdrawn from the release medium and replaced with 1 mL of fresh PBS. The samples were then analyzed via the HPLC method.

The in vitro drug release was analyzed using a Pur-A-Lyzer Mini Dialysis Kit with 3.5 kDa as the molecular weight cut off (MWCO) (Sigma-Aldrich, St. Louis, MO, USA). The release was conducted in a shaking incubator at 100 rpm and 37 °C. The transfersomes containing 0.1 mg R-carvedilol or the same amount of free drug dissolved in PEG 400 (volume 2.5 mL) were added into the dialysis tubes. PBS was used as the release medium and the tubes were immersed in 100 mL of PBS (pH 7.4). At various time intervals (0.5, 1, 2, 3, 4, 6, 8, and 24 h), 1 mL samples were withdrawn from the release medium and replaced with 1 mL of fresh PBS. The samples were analyzed via HPLC.