Rabbit anti met monoclonal antibody

GC cell lines were harvested, lysed by RIPA Lysis Buffer (Solarbio, China), then boiled to prepare for protein. After electrophoresis and PVDF membrane transfer, the target PVDF membrane was blocked, incubated with primary antibody Rabbit anti-Met monoclonal antibody (Abcam, USA) or Rabbit anti-GAPDH Polyclonal antibody (proteintech, China) and secondary antibody HRP-conjugated Goat Anti-Rabbit IgG (H + L) (proteintech, China), and then exposed by electrochemiluminescence (ECL) (Solarbio, China).

With the approval of the Ethics Committee of Lanzhou University Second Hospital and the informed consent of patients, paraffin sections of GC and adjacent tissues from 50 patients with GC who were operated in Lanzhou University Second Hospital were obtained from the Department of Pathology. The steps of IHC are as follows: Following deparaffinization, rehydration, and antigen repair, sections were exposed in 3% H₂O₂ to eliminate endogenous peroxidase activity. Blocked in 3% bovine serum albumin (BSA) for 30 min at room temperature (RT), Rabbit anti-Met monoclonal antibody (Abcam, USA) was incubated as primary antibody overnight at 4 °C, followed by HRP-labeled Goat anti-Rabbit IgG (H + L) (Beyotime, China) as secondary antibody for 1 hour at RT. It is then colored by DAB and counterstained by hematoxylin. The average optical density (AOD) of IHC images was quantitatively analyzed by ImageJ software. The results of IHC were determined by a professional pathologist. The staining intensity was divided into 0 (non), 1+ (weak), 2+ (medium) and 3+ (strong). The staining area was divided into 0 (non), 1+ (<25%), 2+ (25%-50%), 3+ (≥50%). H-score = staining intensity × staining area.