Alkaline phosphatase conjugated goat anti dig fab fragments

Testes were fixed in 4% PFA in PBS overnight at 4 °C, immersed in 30% sucrose, and embedded in O.C.T compound. Sections of 7 μm were cut. Sense and antisense DIG labeled RNA probes were transcribed from a linearized plasmid containing a Tdrd5 cDNA fragment (nucleotides 2160–2676, GenBank NM_001134741.1) using DIG RNA Labeling Mix (Roche). The probes were denatured for 10 min in hybridization cocktail solution (Amresco) and added to the sections for incubation at 65 °C overnight. Sections were then washed, blocked, and incubated with alkaline phosphatase-conjugated goat anti-DIG Fab fragments (Roche) at 4 °C overnight. The positive signal was visualized by BM Purple (Roche).

Testes were fixed in 4% PFA in PBS overnight at 4 °C, after immersed in 30% sucrose, testes were embedded in O.C.T compound, and 7 μm sections were cut. Sense and antisense DIG-labeled RNA probes were transcribed using DIG RNA Labeling Kit (Roche) from a linearized plasmid containing a full length of LINE1 ORF1 (nucleotides 1741–2814, GenBank: M13002.1). After denaturing the probes for 10 min in hybridization cocktail solution (Amresco), the probes were added to the sections and incubated overnight at 65 °C. After washing and blocking, sections were incubated with alkaline phosphatase conjugated goat anti-DIG Fab fragments (Roche) overnight. The positive signal was visualized by adding BM Purple (Roche).