Table 3 Primary antibodies used for immunohistochemistry and their respective pre-treatments, secondary antibodies and chromogens.
From: Unexpected high frequency of neurofibroma in the celiac ganglion of German cattle
Primary antibody | Dilution | Pre-treatments (antigen retrieval/blocking) | Secondary antibody | Dilution | Chromogen |
|---|---|---|---|---|---|
S-100 (Z0311, Dako, Agilent, Waldbronn) | 1:400 | Citrate buffer in microwave/0.2% caseine | Anti-rabbit alkaline phosphatase (AP)-coupled (D0487, Dako, Agilent, Waldbronn) | 1:50 | New Fuchsine |
Vimentin (M7020, Dako, Agilent, Waldbronn) | 1:300 | Protease 0.5% 30 min/0.2% casein, 3% H2O2 | Anti-mouse biotinylated (RPN1001, GE Healthcare Europe GmbH, Freiburg) | 1:100 | AEC |
GFAP (Z0334, Dako, Agilent, Waldbronn) | 1:1000 | −/0.2% casein | Anti-rabbit AP-coupled (see above) | 1:50 | New Fuchsine |
Neurofilament (M0762, Dako, Agilent, Waldbronn) | 1:100 | −/0.2% casein | Anti-mouse alkaline phosphatase (AP)-coupled (D0486, Dako, Agilent, Waldbronn) | 1:500 | New Fuchsine |
PGP 9.5 (PgP9.5-L-CE, Leica Biosystems, Nussloch) | 1:20 | Citrate buffer in microwave/0.2% casein, 3% H2O2 | Anti-mouse biotinylated (see above) | 1:100 | AEC |
MBP (A0623, Dako, Agilent, Waldbronn) | 1:500, overnight 4 °C | −/3% FCS in TBS, 0.1% H2O2 in methanol 30 min | Biotinylated anti-rabbit (RPN1004, GE Healthcare Europe GmbH, Freiburg) | 1:1000 30 min | Tertiary antibody: Strepatvidin—HRP (SN1004, Invitrogen, Thermo Fisher Scientific, USA) chromogen: DAB |
Neurofibromin 1 (LS-B8110/49155, Biozol Diagnostica Vertrieb GmbH, Eching) | 1:50, overnight 4 °C | HCl (4 mM) in steamer/0.2% casein, 3% H2O2 | Biotinylated anti-rabbit (see above) | 1:100 | AEC |
Neurofibromin 2 (sc-331, Santa Cruz Biotechnology, Inc., Heidelberg) | 1:50, overnight 4 °C | HCl (4 mM) in steamer/0.2% casein/3% H2O2 | Biotinylated anti-rabbit (see above) | 1:100 | AEC |