Horseradish peroxidase hrp conjugated anti goat secondary antibody

Testicular tissue sections were dewaxed, rehydrated, and washed 3 times for 5 minutes phosphate-buffered saline (PBS) (Gibco, Grand Island, USA). After slides were microwaved for 20 min and allowed to cool for 1 h at room temperature, endogenous peroxidase activity was blocked in all sections by incubating the sections in 3% H₂O₂ for 15 minutes. The sections were incubated with rabbit anti-mouse Nrf2 and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) antibody (Cell Signaling Technology, USA) overnight at 4 °C. Next day, the slides were washed and incubated with a horseradish peroxidase (HRP)-conjugated anti-goat secondary antibody (Cell Signaling Technology, USA) at a 1:100 dilution for 1 hour. After stained by DAB, the sections were observed under light microscopy.

Immunohistochemistry was performed on paraffin testicle sections at 5 μm of thickness. For specific staining, we incubated slides with rabbit anti-mouse Nrf2 and NF-κB specific antibody (Cell Signaling Technology, USA) overnight at 4 °C. Washing and incubating the slides with a 1:100 dilution horseradish peroxidase (HRP)-conjugated anti-goat secondary antibody (Cell Signaling Technology, USA) for 1 hour the next day.

The tissue sections were dewaxed, rehydrated, and washed 3 times for 5 min phosphate-buffered saline (PBS). After slides were microwaved for 20 min and allowed to cool for 1 h at room temperature, endogenous peroxidase activity was blocked in all sections by incubating the sections in 3% H₂O₂ for 15 min. The sections were incubated with Anti-NLRP3 and Anti-caspase-1 monoclonal antibodies (Cell Signaling Technology, USA) overnight at 4 °C. Next day, the slides were washed and incubated with a horseradish peroxidase (HRP)-conjugated anti-goat secondary antibody (Cell Signaling Technology, USA) at a 1:100 dilution for 1 h. After stained by DAB, the sections were observed under light microscopy.