Keygen one step ihc assay

To establish the nude mouse xenograft model, Bel-7404 cells (1×10⁷cells, in 0.1ml PBS, pH7.4) were subcutaneously injected into the left armpit of a athymic BALB/c nude mouse (SLAC, Shanghai, China). Two weeks later, after xenografts were visible, the mice were treated with Huaier (100ul of solution containing 50mg) or normal saline. Then, all the mice were sacrificed after treatment for consecutive 14 days. Next, tumor xenografts were fixed in paraformaldehyde for 12h and embedded in paraffin. After that, KeyGEN One-Step IHC Assay (KeyGen BioTECH: KGOS60) were used to stain 5um thickness of paraffin sections, and the paraffin sections were incubated with anti-YAP1 (abcam, ab52771). Taking a picture of these paraffin sections. All mouse experiments were performed according to the Interdisciplinary Principles and Guidelines for the Use of Animals in Research, Testing, and Education by the New York Academy of Sciences, Ad Hoc Animal Research Committee.