Sabc immunohistochemical staining kit

The pulmonary tissue sections were incubated at 60 °C for 4 h and immersed in xylene twice for 20 min each, rehydrated by means of a graded series of ethanol, and then incubated with fresh 0.3% hydrogen peroxide in 100% methanol for 30 min at room temperature. The sections were then washed three times using PBS for 15 min. Antigen retrieval was performed with the ImmunoSaver antigen retriever system (Boster, AR0023, USA) at 98–100 °C for 30 min, and sections were cooled to room temperature. Sections were then washed by PBS. Nonspecific binging sites were blocked by incubation with BSA solution (Boster, SA1022, USA) for 30 min. The sections were then incubated with SFTPC antibody (Abclonal, A1835, USA) or Nanog antibody (Thermo Fisher Science, PA1-097, China) in a 1:100 dilution overnight at 4 °C and incubated with secondary antibodies at room temperature for 30 min. The sections were performed according to the steps of the SABC immunohistochemical staining kit (Boster, SA1022, USA). The sections were imaged using a × 40 objective.

These were the tools used: GABA polyclonal antibody (ab8891, Abcam, UK), immunohistochemistry pen (CIRISC, Japan), SABC immunohistochemical staining kit, DAB color kit (Boster Biological Technology, Ltd., Wuhan, China), Leica slicer (RM 2235, Leica, Germany); dynamic plantar aesthesiometer (37450, Ugo Basile, Italy), culture inverted microscope (TS100-F, Nikon, Japan), BX51 optical microscope system (OLYMPUS, Japan), and Visopharm stereology image system (Denmark).

The harvested right testis was weighed and grounded in liquid nitrogen. All grounded tissues from the infected mice were stored at −80°C until virus titration. Viral RNAs were extracted with a universal RNA kit [Cofitt Life Sciences (HK) Limited] and determined by real-time qRT-PCR on a Roche LightCycler® 96 using TB Green Premix Ex Taq ^TMII (TaKaRa, Japan). After comparison with a standard curve, viral load in testicles was expressed as viral RNA copies per μg RNA. The primer sets for these assays are described (Table 1).
The paraffin blocks of tissues were used for an immunohistochemical assay by the method of Wenqiang Ma and Wanpeng Yu (18 (link), 19 (link)) by a SABC immunohistochemical staining kit (BOSTER, SA1020). Deparaffinization, rehydration, and antigen retrieval were performed on the 5-mm paraffin sections of the testis or epididymis. The tissue sections were treated with 3% H2O2 in PBS (pH7.6) for 10 min and 5% BSA for 30 min. And then we incubated the sections at 4°C overnight with a Rabbit antibody against the E2 protein of the Getah virus as primary antibodies. The sections were rinsed with PBS three times and incubated with biotin-labeled goat anti-rabbit as secondary antibodies for 30 min. After incubation with SABC for 30 min, the specific binding on the sections was visualized by DAB. The sections were counterstained with hematoxylin.