Maxvision tm hrp polymer anti mouse ihc kit

Paraffin-embedded sections of placental tissues were prepared, heated at 65℃ for 2 h, and dewaxed in xylene I and II. Afterward, sections were treated with 100, 95, 85, and 70% ethanol, immersed for 2 min in boiled 0.01 M citrate buffer, cooled for 30 min and subsequently rinsed with phosphate buffered saline. Subsequently, sections were cultivated for 20 min in 3% H₂O₂ in order to block the endogenous peroxidase activity. Next, sections were blocked by normal goat serum and cultivated at 4℃ overnight with primary antibodies (CXCL12, 1:100; CXCR4, 1:100; CXCR7, 1:200; all from R&D). Lastly, sections were subjected to incubation with MaxVision^TM HRP-Polymer anti-mouse IHC Kit (Maixin, Fuzhou, China), DAB development, alcohol gradient dehydration, xylene immersing, neutral gum blocking, as well as observation under a microscope. A double-blind method was utilized to judge each section independently. The images were collected and saved, the immunohistochemical results were analyzed using Image-Pro-Plus image analysis software (Media Cybernetics, Bethesda, MD, USA), and the integrated optical density (IOD) values were calculated. IHC results were interpreted according to the following criteria: when the cell membrane or cytoplasm showed brownish-yellow particles, the cells were positive.