Rhodamine red x affinipure goat anti rabbit igg h l

The cells were fixed with 4% paraformaldehyde at room temperature for 15 min and subsequently permeabilized in 0.2% Triton X-100 (Sigma-Aldrich X100) in PBS for 5 min. The cells were blocked with PBS containing 5% normal goat serum and 0.1% Triton X-100 for 20 min. The cells were incubated with primary antibody (Tom20-specific antibody, Santa Cruz Biotech. SC-11415) at 4 °C for overnight. After washing with PBS containing 0.1% Triton X-100, the cells were incubated with secondary antibody (Rhodamine Red-X AffiniPure Goat Anti-Rabbit IgG (H+L), Jackson ImmunoResearch Laboratories 111-295-144) for 1 h. DAPI Fluoromount-G Mounting Medium (Southern Biotech 0100-20) was used to counterstain the nuclei. All images were obtained with a fluorescence microscope (Olympus BX53, Shinjuku, Tokyo, Japan).

10x PBS recipe: https://www.protocolsonline.com/recipes/phosphate-buffered-saline-pbs/. Reagents for PBST: Triton X-100 at Sigma-Aldrich, St. Louis, MO, USA (CAS# 9002-93-1), BSA (albumin fraction V) at Chem-Impex-International, Wood Dale, IL, USA (CAS# 9048-46-8), Ana2, and Sas-6 antibodies were kindly provided from Jordan Raff laboratory. Gamma tubulin antibodies (GTU88) at Sigma-Aldrich (Catalog No. T6557) Cy™5 AffiniPure Goat Anti-Rabbit IgG (H+L) at Jackson Immuno Research Laboratory, Philadelphia, PA, USA (Code: 711-175-152), Rhodamine Red-X-AffiniPure Goat Anti-Rabbit IgG (H+L) at Jackson Immuno Research (Code: 111-295-144), Alexa Fluor 647 at Jackson Immuno Research (Code 711-606-152), and Poc1 N-terminus antibody was made within our lab and is available upon request. Glutaraldehyde at Electron Microscopy Sciences (CAS #111-30-8). Cacodylate buffer was kindly provided by Dr. William Gunning at the University of Toledo. EMBed 812 resin at Electron Microscopy Sciences (CAS #13940). Reynolds Lead citrate at Electron Microscopy Sciences (CAS #10099-74-8). ImageJ software is available at https://imagej.nih.gov/ij/.