Hrp labeled goat anti mouse secondary antibody

Fresh skin samples of Inner Mongolia Cashmere goat were obtained during catagen, telogen, and anagen and fixed in 4% paraformaldehyde for 24 h. Following dehydration in pure alcohol, the sections were placed in benzene to replace ethanol prior to paraffin-embedding. The slices of tissue samples (8-μm-thick) were incubated with xylene for dewaxing, subjected to gradient alcohol hydration, and incubated at room temperature in 3% hydrogen peroxide (H₂O₂) to inactivate the endogenous catalase. Next, the specimens were flushed with phosphate-buffered saline (PBS), antigen retrieval was carried out in citrate buffer, and blocking with 5% BSA at room temperature for 1 h. Subsequently, the samples were incubated overnight at 4 °C with mouse monoclonal anti-vimentin antibody (Abcam. 1:250); 5% albumin from bovine serum (BSA) was used as a negative control. Then, the samples were washed with PBS and incubated with the HRP-labeled goat anti-mouse secondary antibody ((Beyotime. 1:500) for 1 h at 37 °C. Finally, the sections were stained with hematoxylin and observed under a microscope.

Tim-AIII (purity >98%) was purchased from Chengdu Herbpurify Co., LTD (Z-019, China) and was dissolved in dimethyl sulfoxide (DMSO, 196055, MP Biomedicals, USA) to prepare a stock solution of 10 mM and stored at -20°C. Mouse anti-GPX4 (67763-1-Ig) antibody, rabbit anti-HSP90 (13171-1-AP), E-Cadherin (20874-1-AP), Vimentin (10366-1-AP), Snail-1 (13099-1-AP), Snail-2 (12129-1-AP), N-Cadherin (22018-1-AP) antibodies and HRP-conjugated anti-heavy chain of rabbit IgG antibody (SA00001-7H) were purchased from Proteintech (Wuhan, China). Rabbit anti-GPX4 (A1933), FTL (A11241), HMOX-1 (A1346), SLC40A1 (A14885), SLC7A11 (A13685) Ubiquitin (A19686) and β-actin (AC004) antibodies were purchased from ABclonal (Wuhan, China). The HRP-labeled goat anti-rabbit IgG and HRP-labeled goat anti-mouse secondary antibody was purchased from Beyotime (Shanghai, China). Chloroquine (CQ, HY-17589A), Z-VAD-FMK (HY-16658B), Necrostatin-1 (Nec-1, HY-15760), Ferrostatin-1 (Fer-1, HY-100579), N-acetylcystein (NAC, HY-B0215), tanespimycin (HY-10211), trolox (HY-101445), cycloheximide (CHX, HY-12320) and MG-132 (HY-13259) were purchased from Med Chem Express (Shanghai, China). The recombinant human HSP90 alpha protein (ab48801) was provided by Abcam (Shanghai, China). The recombinant Human GPX4 protein (P0633) was provided by Fine Biotech (Wuhan, China).