Bis tris sds page gel electrophoresis

The quantitative western blot analysis was used to detect sPDGFRβ in human CSF (ng/mL), as we previously reported.²⁶, ²⁷ Briefly, CSF samples and recombinant human PDGFRβ carrier‐free protein (Cat. No. 385‐PR‐100/CF, R&D Systems) were subjected to 4–12% Bis‐Tris SDS/PAGE gel electrophoresis (Thermo Scientific) for 2 h at 150 V and subsequently transferred to a nitrocellulose membrane. The membrane was blocked for 1 h with a superblock blocking buffer (Cat. No. 37537, Thermo Scientific). The membrane was incubated with a primary antibody for PDGFRβ (Cat. No. AF1042, R&D Systems) overnight, then incubated with donkey anti‐goat IgG secondary antibody (Cat. No. A15999, 1:5000 dilution, Thermo Scientific) for 1 h at room temperature, treated with Western ECL detection solution (Cat. No. 34075, Thermo Scientific), exposed to CL‐Xposure film (Cat. No. 34091, Thermo Scientific), and developed in an X‐Omat 3000 RA film processor (Kodak). Images were acquired, and densitometry analysis was performed using NIH Image J software.