Anti cd68 antibody

The liver sections from the right lobe and mesenteric tissues were fixed in 10% formalin buffer (pH 7.4) and embedded in a paraffin block. Hematoxylin–eosin (H&E), Masson, and Sirius Red staining were used on the liver sections, followed by random evaluation under a light microscope by an expert pathologist.
For IHC staining, liver sections were incubated with antimatrix metalloproteinase-2 (MMP-2) antibody (1:150, Servicebio, China), antimatrix metalloproteinase-9 (MMP-9) antibody (1:300, Servicebio, China), antivascular endothelial growth factor (VEGF) A antibody (1:250, Servicebio, China), antivon Willebrand factor (vWF) antibody (1:1,000, Servicebio, China), anticytokeratin 19 (CK-19) antibody (1:1,000, Servicebio, China), anti-sEH antibody (1:50, Absin, China), anti-CD31 antibody (1:300, Servicebio, China), and anti-CD68 antibody (1:150, Servicebio, China), overnight at 4°C with phosphate-buffered saline (PBS) as negative control. Subsequently, the sections were incubated with appropriate HRP-conjugated goat antirabbit secondary antibody for 60 min, followed by restaining with hematoxylin. The collagen deposition volume and stained area were calculated with IHC Profiler plugin in ImageJ (version 1.53, USA). The results were expressed as the proportions of the stained areas. The total area and average values were taken from five rats in each group.

We obtained the tissue microarray from the Outdo Biotech company (HOrg-C110PT-01, Shanghai, China) and the ethics was approved. Briefly, the paraffin sections of pan-cancer samples were deparaffinized and then were blocked with 3% H2O2 and 2% BSA after antigen retrieval. Anti-PDIA5 antibody (Rabbit, 1:100, Proteintech, China), anti-CD68 antibody (Rabbit, 1:3000, Servicebio, China), and anti-CD163 antibody (Rabbit, 1:3000, Proteintech, China) were sequentially applied, followed by horseradish peroxidase-conjugated secondary antibody (GB23301 and GB23303; Servicebio, China) incubation and tyramide signal amplification (TSA) (CY5-TSA, CY3-TSA, and FITC-TSA; Servicebio, China) incubation. And then, 4’,6-Diamidino2-phenylindole dihydrochloride (DAPI) was applied for the nuclei staining. The stained slides were visualized for multispectral images under the Pannoramic Scanner (3D HISTECH, Hungary). DAPI glows blue by UV excitation wavelength 330-380nm and emission wavelength 420nm in the fluorescence spectra. CY5, FITC, and CY3 glow pink, green, and red. The excitation wavelength was 608-648nm, 465-495nm, and 510-560nm, respectively, with an emission wavelength of 672-712nm, 515-555nm, and 590nm.