Peroxidase labeled anti dig antibody

Method of preparing paraffin sections of hen prehierarchical follicles and further histological processing were as our previously described [30 (link)]. After the sections were mounted on slides, a modified H&E staining procedure was used, as described by Zheng (2005) [31 ]. The sections were examined using a JNOEC XS-213 biological microscope (Jiangnan Optics & Electronics Co., Ltd. Nanjing, China) at magnifications of ×10, ×40 and ×200.
Frozen ovarian tissues were serially sectioned at a thickness of 10 μm on a cryostat for in-situ hybridization (ISH) experiments, using Digoxin (DIG)-labeled cRNA probes corresponding to the genes listed in Table 1, and their sense sequence probes against the genes were used to confirm the specificity of the binding. The DIG-labeled in-situ hybridization was performed as previously described with slight modifications [30 (link)]. mRNA expression of the candidate gene was then detected with a peroxidase-labeled anti-DIG antibody (dilution, 1:1000; Roche Diagnostics, Indianapolis, IN, USA) and chromogenically developed by incubating the slides with DAB staining solution (containing 3, 3’-diaminobenzidine 50 mg, 0.05 M Tris buffer (TB; 0.05 M Tris-hydrochloride buffer, pH 7.5) for 5 min at room temperature, and the sections were then counterstained with hematoxylin for morphological observation.