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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