Genital tract lesions in sexually mature Göttingen minipigs during the initial stages of experimental vaginal infection with Chlamydia trachomatis serovar D

Table 1 Details of immunohistochemical protocols

Target antigen	Manufacturer	Dilution and incubation	Detection method and chromogen
Mouse anti-Chlamydia^a	Progen Clone Acl-C	1:100 o/n RT	Vectastain Elite-ABC - Mouse
Mouse anti-pig CD3e^b	Southern Biotech SB 4510-01	1:1000 o/n 4 °C	Ultravision One DAB
Goat anti-pig IgM^c	Nordic Biosite Lot no: A100-100A-11	1:5000 1 h RT	Vectastain Elite-ABC – Goat AEC
Goat anti-pig IgA^c	Nordic Biosite Lot no: A100-102A-16	1:4000 1 h RT	Vectastain Elite-ABC – Goat AEC
Goat anti-pig IgG-Fc^c	Nordic Biosite Lot no: A100-104A-12	1: 7000 1 h RT	Vectastain Elite-ABC – Goat AEC
Rabbit anti-mouse/rat Cox-2^d	Cayman chemicals Lot. no: 0428356-1	1:400 o/n 4 °C	Ultravision One AEC
Mouse anti-sheep IL-8^e	Abd Serotec Clone 8 M6, MCA 1660	1:1000 o/n 4 °C	Ultravision LP AEC

For antigen retrieval, the following procedures were used: boiling in ^aCitrate buffer pH 6.0 for 2 × 5 min ^bTEG buffer pH 9.0 for 2 × 5 min, ^cCitrate buffer pH 6.0 for 5 min ^dTED buffer pH 9.0 2 × 5 min. For ^eIL-8, a DIVA decloaker solution in a pressure cooker (2100 retriever, Biocare Medical) was used. Blocking for endogenous peroxidase was carried out using a solution of hydrogen peroxide (0.6 %) for 15 min for all stainings except Chlamydia, where a 1 % dilution for 30 min was used

ISSN: 1746-6148