Rabbit igg hrp conjugated antibody

The following primary antibodies were utilized: (1) goat polyclonal anti-rat CD31 antibody (R&D Systems, American), dilution 1:250; (2) rabbit polyclonal anti-rat VEGF and VEGFR2 antibody (BiossAntibodies, Beijing, P.R. China), dilution 1:250; (3) rabbit polyclonal anti-rat MMP2 and MMP9 antibody (Bioss Antibodies, Beijing, P.R. China), dilution 1:250, (4) rabbit polyclonal anti-rat ET-1 antibody (Bioss Antibodies, Beijing, P.R. China), dilution 1:200. The goat IgG HRP-conjugated antibody and rabbit IgG HRP-conjugated antibody were purchased from R&D. Sections were finally stained with 3ʹ-diaminobenzidine (DAB). All steps were performed strictly according to the manufacturer’s instructions. Images were acquired using an Olympus BX51 clinical microscope, DP70 digital camera and software (Olympus). The images were analyzed using Image-Pro Plus (Media Cybernetics). Microvessel density (MVD) was measured in terms of the CD31-positive endothelial areas. Every positive area was calculated as one microvessel.18 (link) The expression of VEGF, VEGFR2, MMP2, MMP9 and ET-1 was measured according to the immunohistochemical mean density= IOD sum/area in three randomly selected microscopic fields for each slide.

The levels of TC (cat. no. CS0005; Sigma-Aldrich; Merck KGaA), TG (cat. no. MAK266; Sigma-Aldrich; Merck KGaA) and LDL-r (cat. no. RAB0707; Sigma-Aldrich; Merck KGaA) were measured using ELISA kits. An Oil Red O staining kit (cat. no. ab150678; Abcam) was also employed. The primary antibodies were anti-β-actin (cat. no. ab8227; 1:4,000; Abcam), anti-SRA1 (1:500; cat. no. sc-166139; Santa Cruz Biotechnology, Inc.), anti-CD36 (1:3,500; cat. no. ab133625; Abcam), anti-SREBP2 (1:3,000; cat. no. ab30682; Abcam) and anti-LDL-r (1:4,000; cat. no. ab52818; Abcam). The secondary antibodies used were Goat IgG horseradish peroxidase (HRP)-conjugated antibody (1:2,000; cat. no. HAF017; R&D Systems, Inc.) and Rabbit IgG HRP-conjugated antibody (1:2,000; cat. no. HAF008; R&D Systems, Inc.).