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[beta]-Actin (42 kDa) was used as loading control, and rabbit polyclonal anti-MTA1 antibody (cat: bs-1412R, dilutions: 1: 1500, Bioss Antibodies, China) or rabbit polyclonal anti-[beta]-actin antibody (cat: bs-0061R, dilutions: 1: 50,000, Bioss Antibodies, China) was used in this assay.
The rabbit anti-murine KEAP1 (primary antibody), the rabbit anti-murine NFE2L2 (primary antibody) antibodies, and the rabbit two-step kit (secondary antibody) were purchased from Bioss (Beijing, People's Republic of China).
For immunohistochemical analysis, sections were blocked with 5% bovine serum albumin (BSA) for 20 min and incubated with antibodies against TNF-[alpha] (1: 150 dilution, Bioss, Beijing, China) and IFN-[gamma] (1: 100 dilution, Bioss, Beijing, China) at 4[degrees]C for 12 h, followed by incubation with goat anti-rabbit immunoglobulin G (ZSGB-BIO, Beijing, China) at 25[degrees]C for 2 h; then the sections were counterstained with hematoxylin.
Further identification and purity of Sertoli cells were assessed by immunocytochemistry (IHC) labeled with antifollicle-stimulating hormone receptor (anti-FSHR) antibody (Bioss Biology Corporation, Beijing, China).
After blocking in 5% nonfat milk diluted with TBST (tris-buffered saline/Tween-20) for 1 h at room temperature, the membranes were separately incubated with rabbit anti-ILF2 monoclonal antibody (1 : 3000; Abcam) and rabbit anti-GAPDH antibody (1 : 3000; Bioss) at 4[degrees]C overnight.
The membrane was incubated overnight at 4[degrees]C with rabbit anti-PGE2 polyclonal antibody (1:5,000) (Bioss Inc., Woburn, MA, USA).
The samples were then incubated with primary antibodies against 5-bromodeoxyuridine (BrdU, Bioss Biotechnology, Beijing, China), runt-related transcription factor 2 (RUNX2, Santa Cruz Biotechnology, CA, USA), osteocalcin (OCN, Abcam plc., Cambridge, UK), osteopontin (OPN, Novus Biologicals, Colorado, USA), osteoprotegerin (OPG, Abcam plc., Cambridge, UK), or receptor activator of the nuclear factor-?B ligand (RANKL, Novus Biologicals, Colorado, USA) at manufacturer-recommended dilutions in a humidified chamber at 4[degrees]C overnight.
(16.) Syed Muhammad, Mukarram Shah, Farhat Ullah, Syed Muhammad, Hassan Shah, Mohammad, Zahoor and Abdul Sadiq .Analysis of chemical constituents and antinociceptive potential of essential oil of Teucrium Stocksianum bioss collected from the North West of Pakistan.
We obtained anti-phycocyanin from Bioss (Woburn, MA), microtiter ELISA plates from Corning (Corning, NY), microcystin LR and anti-FceRI from EMD Millipore Corporation (Billerica, MA), Eagle's Minimum Essential Medium (EMEM) from ATCC (Manassas, VA), and phycocyanin, G418, and pNPP substrate from Sigma-Aldrich (St.
Anti-HMGB1 antibody (ab79823), anti-TGF-[beta]1 antibody (ab64715) and anti-type I collagen antibody (ab34710) were Abeam products, while anti-a-smooth muscle actin ([alpha]-SMA) antibody (bs10196R) was purchased from Bioss and anti-[beta]-actin antibody (E1A7018) was from EnoGene.
Hunte and her team conducted the research at the Institute for Biochemistry and Molecular Biology of the Faculty of Medicine and the Centre for Biological Signalling Studies BIOSS at the University of Freiburg.