Anti pdgf bb

Protein contents of VEGF-A, PDGF-BB, IL-1β, and TNF-α in ulcer tissue homogenates were evaluated by western blot. In brief, the protein concentrations of homogenates were determined using a BCA protein assay (Pierce Biotechnology, Rockford, IL, USA). Equivalent amount of protein samples (30 μg) was separated on an SDS polyacrylamide gel and transferred onto a polyvinylidene difluoride membrane (Millipore). After blocking in TBS containing 5% nonfat milk for 2 h at room temperature, the membranes were incubated overnight with 1 : 1,000 diluted anti-VEGF-A (Abcam), 1 : 2000 diluted anti-PDGF-BB (Abcam), 1 : 200 diluted anti-IL-1β (Santa Cruz Biotechnology), 1 : 500 diluted anti-TNF-α (Abcam), and 1 : 8000 diluted anti-β-actin (Tianjin Sungene Biotech Co., Ltd.), respectively. Binding of the primary antibody was detected using a HRP-conjugated secondary antibody (Tianjin Sungene Biotech Co., Ltd.). Positive bands were visualized using an ECL kit (Bio-Rad) and then captured on X-ray film. Housekeeping protein β-actin was used as a loading control. The density of each band was quantified using Quantity One software (Bio-Rad Laboratory) and normalized to their respective control.

Ginsenoside Rb1 (purity > 99.5%) was purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). SSD cream (1%) was used as a positive control.Anti-FGF-2, anti-PDGFR-β, anti-PDGF-BB and anti-β-actin antibodies were purchased from Abcam (Shanghai, China). A BCA protein quantification kit was purchased from BiyunTian (Shanghai, China). Electrochemiluminescence (ECL) reagent was purchased from Millipore (Billerica, MA, USA). A total RNA extraction kit was purchased from Tiangen (Beijing, China). A RevertAid First Strand cDNA Synthesis Kit was purchased from Thermo Fisher Scientific (Waltham, MA, USA). RT-PCR Master Mix was purchased from Vazyme (Nanjing, China).The various other chemicals used were of analytical grade and commercial origin.