Anti vδ2 tcr primary antibody

Tissues were harvested and fixed in Formalde-Fresh Solution overnight at 4°C, washed with 1X PBS and transferred to 70% ethanol before paraffin embedding and sectioning. Tissue embedding and sectioning were performed by the Histotechnology Facility (The Wistar Institute). For immunohistochemistry (IHC) studies, tissue sections were deparaffinized in xylene, rehydrated in ethanol (100%-95%-80%-70%) and distilled water. The endogenous peroxidase activity was quenched with 0.5% hydrogen peroxide in methanol for 10 minutes. The slides were washed under tap water for 5 minutes, simmered in Tris-EDTA buffer, washed with PBS before blocking them in 5% BSA blocking solution for 1 hr. The tissue sections were subsequently incubated with anti Vδ2-TCR primary antibody (Bio Legend cat #331402) and anti-E. coli antibody (Abcam ab137967) 1:50 in 5% BSA overnight at 4°C, washed next day with 1X PBS and incubated with AF647 (Invitrogen, cat # A21236) and AF488 (Bio Legend Cat# 406416) secondary antibodies 1:200 for 45 min. DAPI (1:5000) was added for 5 minutes and the samples mounted using Cytoseal 60 or Mounting Medium (Electron Microscopy Sciences). Specimens were photographed using 80i upright microscope and analyzed with the NIS-Elements Basic Research software.