Table 1 Immunohistochemical protocols used for cellular type characterization
Primary antibody (Ab) | Specifity | Dilution Ab | Epitope demasking | Secondary Ab |
|---|---|---|---|---|
CD3 (Novocastra-CL-L-CD3–565), mouse monoclonal | Pan T cell marker | 1:500 in TBS 1% | Microwave in citrate pH 6 20 min | Anti-mouse biotinylated (1:200) |
CD20 (ThermoFisher.-PA516701), rabbit polyclonal | Pan B cell marker | 1:200 in TBS + BSA 1% | Microwave in citrate pH 6 20 min | Anti-rabbit biotinylated (1:200) |
Iba1 (WAKO 019_19741), rabbit polyclonal | Macrophages | 1:1000 in TBS + BSA 1% | Los Angeles pH 9 40 min 95 °C | Anti-rabbit biotinylated (1:200) |
Lambda (Dako A0193), rabbit polyclonal | Plasma cells | 1:1000 in TBS + BSA 1% | Triton 1% 20 min room temperature | Anti-rabbit biotinylated (1:200) |