Df7766

Quantitative IHC is a technique used to measure the density of protein expression in immunohistochemically stained images. This method involves calculating the chromogenic density or color and the associated signal intensity to obtain a quantitative result that can be used to study the amount or trend of protein expression in a cell or tissue. The cervical spinal cord of each group was collected and fixed in 4% paraformaldehyde for 3 days at 4°C. Fixed samples were embedded in paraffin, and 5μm-thick sections were obtained. Slices were deparaffinized in xylene and rehydrated in graded alcohol solutions. The tissue sections were repaired in 3% citric acid repair solution under high pressure and then incubated in 3% H₂O₂ for 10 min. Next, the sections were washed with distilled water for 5 min and soaked with phosphate-buffered saline (PBS) for 1min. The sections were then incubated with primary antibody to PSD-95 (AF7839, 1:100, Affinity), GAP-43 (DF7766, 1:90, Affinity) at 4°C overnight. The sections were washed three times with PBS buffer for 5 min on the next day, incubated with the secondary antibody at 37°C for 20 min, followed by reaction termination using 3,3’-diaminobenzidine (DAB). BX43 microscope (Olympus, Japan) was used to capture the immunohistochemistry images of the slices, and Image-Pro Plus software was used to quantify the captured image.