Ihc optimized protocol

The expression of platelet-derived growth factor-C (PDGF-C) in the rat brain’s hippocampus tissue was determined using an immunohistochemistry (IHC) analysis. A total of 2–3 mm thick coronal sections were cut throughout the dorsal hippocampus. Brain sections were processed in a Thermo tissue processor, according to the manufacturer’s protocol for 16 h, this included fixation, dehydration, clearing, and wax infiltration. Brain tissues sections were embedded in molten paraffin wax (Thermo, Waltham, MA, USA) and cooled out; the formed paraffin wax mould was then trimmed for obtaining 3 um sections using a Leica microtome, and the sections were fished out using Poly-Lysin coated slides (Thermo) for IHC staining. All the prepared slides were left at room temperature overnight for dehydration. The IHC-optimized protocol (Abcam) was followed. The primary antibody (Rabbit Polyclonal against PDGF-C, Abbexa) and secondary antibody (Rabbit specific HRP_DAB IHC Detection Kit—Micro-polymer v2d, ab236469, Abcam) were used. The method was performed according to the manufacturer’s (Abcam) instructions. No primary antibody control slide for each sample was run in parallel with the tested slide. The slides were observed under the microscope to score the intensity of staining.

The IHC study was performed by Eman Biodiscoveries Sdn. Bhd., Sungai Petani, Kedah, Malaysia. IHC optimized protocol (Abcam, USA) was followed. The primary antibody (Rabbit Polyclonal against Platelet-Derived Growth Factor C (PDGFC), Abbexa-UK) and secondary antibody (Rabbit specific HRP_DAB IHC Detection Kit—Micro-polymer v2d, ab236469, Abcam, USA) was used. The method was performed according to manufacturer (Abcam) instructions. The slides were observed under the microscope for identifying the intensity of immunostaining in the hippocampus for PDGF-C expression.