Sabc rabbit igg pod kit

IHC staining was performed on 8 μm frozen sections of the aortic root to detect the proportion of macrophages, endothelial cells, and SMCs. Frozen sections were air-dried, fixed with cold acetone, and incubated in anti-CD31 antibody (1:50 dilution; ab28364, Abcam, USA), anti-CD68 antibody (1:50 dilution; ab125212, Abcam, USA), and anti-alpha smooth muscle actin antibody (1:200 dilution; ab5694, Abcam, USA), respectively, at 4 °C overnight. Substitution of PBS for the specific primary antibody was performed as a negative control. All steps were operated following the instruction manual of the SABC (rabbit IgG)-POD kit (Solarbio, Beijing, China). Finally, slides were stained with DAB, counterstained with hematoxylin, dehydrated, and mounted. Images were detected using a Nikon A1R confocal laser scanning microscope (Nikon, Tokyo, Japan). Image Pro-Plus 6.0 software (Media Cybernetics, Rockville, USA) was used to analyze the positively stained areas. At least three sections were analyzed per aortic root.

Mouse insulin (EMD Millipore), TNF-α and IL-6 (Abcam) ELISA kits, anti-DNA (cat. no. AC30-10) monoclonal antibody (Novus Biologicals, Ltd.), D-Luciferin, sodium salt D-Fluorescein sodium salt (cat. no. 40901ES01; Shanghai Yeasen), LUC-labeled lentiviral particles (cat. No. GM-0220PC, Shanghai; Genomeditech), RIPA lysis buffer, nitrocellulose membranes, and an SABC (rabbit IgG)-POD kit were obtained from Beijing Solarbio Science & Technology Co., Ltd. A blood glucose meter and test strips were acquired from ACCU-CHEK (Accu-ChekSoftclix;). TRIzol® was purchased from Invitrogen (Thermo Fisher Scientific, Inc.). Antibodies against PI3K, P85, phospho-PI3K P85 (cat. no. Y607), insulin receptor β (IR), p-IR (cat. no. Y1185), PTEN, p-PTEN (cat. no. S380+T382+T383), glucose transporter 4 (cat. no. GLUT4), and IGF-1R were obtained from Abcam. Antibodies against Akt, p-Akt (ser 473), ERK, and p-ERK (1/2) (Thr202/Tyr204) were obtained from Cell Signaling Technology, Inc. Anti-GAPDH, anti-mouse IgG, and anti-rabbit IgG were obtained from Cell Signaling Technology, Inc. Other reagents used were of analytical grade.