Alexa fluor 647 donkey anti rat igg

Tissue sections were analyzed by chromogenic IHC targeting mCherry and counterstained with hematoxylin. All staining was performed by HistoWiz Inc using the Leica Bond RX Automated Stainer (Leica Microsystems, Wetzlar, Germany). IF analysis was performed as previously described (19 (link)). Briefly, sections were blocked with 3% bovine serum albumin, 2% donkey serum, and 0.5% Triton X-100 (Sigma-Aldrich, St. Louis, MO, USA) for 1 hr at RT. The tissue sections were then incubated overnight at 4°C with primary antibodies, followed by incubation with the appropriate secondary antibodies for 1 hr at RT. Sections were mounted with an antifade solution containing 4’,6-diamidino-2-phenylindole (DAPI) (Vector Laboratories, Newark, CA, USA) to stain the nuclear DNA. Antibodies from Thermo Fisher Scientific were used: Alexa Fluor 488 phalloidin (1:200), Alexa Fluor 488 donkey anti-mouse IgG (1:1000), and Alexa Fluor 647 donkey anti-rat IgG (1:1000). Antilaminin gamma 1 (A5, Novus Biologicals, Centennial, CO, USA) was used at a 1:400 dilution.