The DNeasy® Mini Procedure uses a spin-column of DNA-binding membrane and a buffer system for cell lysis, DNA binding and elution [7].
Total protein (20 μg per sample) was separated on a 8%T/3%C, 6M urea, SDS-polyacrylamide gel using a Tris/glycine buffer system, and transferred to a PVDF membrane overnight.
30 µg of total RNA was resolved in 15% of denaturing polyacrylamide gel containing 7 M urea in 0.5XTBE buffer system and transferred onto Zeta-Probe membrane (BioRad) in 0.5XTBE.
Approximately 50 μg of protein/lane were loaded and run on the polyacrylamide gel with a Tris/glycine running buffer system and then transferred onto a PVDF membrane.
Samples (20 μg) were separated by 13.5% (v/v) SDS−PAGE using the Tris-tricine buffer system and proteins were transferred onto polyvinylidene difluoride membrane (Millipore).
For Western blot analysis, 25 μg of protein was resolved on 8% bis-tris gels in 3- N-morpholino propanesulfonic acid buffer system and electrotransferred onto a 0.45 μm nitrocellulose membrane.
Cell lysates were separated by SDS-PAGE on a 7.5% polyacrylamide gel in a discontinuous buffer system and gels were blotted on nitrocellulose membranes (Hybond C, GE Healthcare Europe GmbH).
Cell lysates were separated by SDS-PAGE and proteins were transferred to polyvinylidene difluoride membranes (Millipore) for 20 min at 150 mA, using a Tris-glycine buffer system.
SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) was performed using a MOPS buffer system (Thermo Fisher Scientific), and gels were subsequently transferred to PVDF membranes using a wet blotting system.
The treatment is performed by putting the patient's blood through the dialyser where it comes into contact with a semipermeable membrane with dialysis fluid, the content of which allows for regeneration of the blood buffer system.
The bacterial lysate was subsequently centrifuged (3,000× g, 30 min, 4°C), passed through a 0.45 µm pore-size membrane filter (Millipore Corporation, Billerica, MA, USA) and adjusted (with HCl or NaOH) to the pH of the buffer system.
