A594 goat anti rabbit igg

Immunofluorescence staining was performed as described previously (11 (link)). In brief, after embedding with a frozen section compound (Leica, #3801480), biopsies were sectioned into 4-μm on a microtome (Leica CM1950). For immunofluorescent staining, the sections were fixed in pre-cooled methanol (-20°C) for 5 minutes, after washing twice with PBS. The sections were then blocked with PBS/5%BSA/Fcγ blocker at 4°C for 1 hour. Primary antibodies were incubated with sections at 4°C overnight. After washing twice with PBS, the sections were incubated with fluorescent-coupled secondary antibodies for 1 hour at room temperature. After two further washes, the sections were mounted and imaged on an immunofluorescence microscope (Leica DMI3000B).
Antibodies used for staining included: Mouse-anti-human CD3 monoclonal antibody (Invitrogen, # MA1-21454), Rabbit-anti-human CTLA4 polyclonal antibody (Invitrogen, # PA5-115060), Rabbit-anti-human PD1 polyclonal antibody (Invitrogen, # PA5-20350), Rabbit-anti-human CCL4 polyclonal antibody (Invitrogen, #PA5-114961), Rabbit-anti-human FGFR2 polyclonal antibody (Invitrogen, #PA5-14651), A594 goat-anti-rabbit IgG (Affinity, #S0006), A488 donkey-anti-mouse IgG (Invitrogen, #A-21202) and DAPI (Invitrogen, #D21490).

Biopsies were taken from six cervical patients with cervical cancer. After embedding with a frozen section compound (Leica, #3801480), biopsies were sectioned into 4-μm on a microtome (Leica CM1950). For immunofluorescent staining, the sections were fixed in pre-cooled methanol (-20°C) for 5 minutes, after washing twice with PBS. The sections were then blocked with PBS/5%BSA/Fcγ blocker at 4°C for 1 hour. Primary antibodies were incubated with sections at 4°C overnight. After washing twice with PBS, the sections were incubated with fluorescent-coupled secondary antibodies for 1 hour at room temperature. After two further washes, the sections were mounted and imaged on an immunofluorescence microscope (Leica DMI3000B).
The antibodies/materials used for immunofluorescent imaging included: A488 mouse-anti-human CD206 (Invitrogen, #MA5-23656), PE mouse-anti-human CD163 (Invitrogen, #12-1639-42), A488 mouse-anti-human EpCam (Abcam, #ab237395), mouse-anti-human KRT16/Cytokeratin Pan Antibody Cocktail (Invitrogen, #MA5-13203), rabbit-anti-human CRCT1 (Invitrogen, #PA5-23539), A594 goat-anti-mouse IgG (Invitrogen, #A11032), A594, goat-anti-rabbit IgG (Affinity, #S0006), rabbit-anti-human DR5 (Invitrogen, # PA1-957), A647 donkey-anti-rabbit IgG (Invitrogen, #A32795) and DAPI (Invitrogen, #D21490).