Anti ahr antibody

Immunochemical staining was performed as described previously [27 (link)]. Cells seeded in 48-well plates were fixed with 4% paraformaldehyde for 15 min and washed three times with PBS. Then, the cells were permeabilized with 0.3% Triton X-100 for 30 min followed by blocking with Immunol Staining Blocking Buffer (Beyotime, Shanghai, China) for 1 h at room temperature. The cells were incubated with anti-AhR antibody (1:100, Proteintech) overnight at 4 °C and rewarmed at 37 °C for 1 h the next day. The cells were incubated with Alexa-488-conjugated secondary antibody (1:500, Proteintech) for 50 min at 37 °C, stained with DAPI (Beyotime, Shanghai, China), washed three times, and viewed by a laser scanning confocal microscope (Olympus, Tokyo, Janpan).

The animal tissues were prepared as described above including fixation, decalcification, tissue embedding, and section. After deparaffinization and rehydration, samples were blocked with serum. RAW 264.7 cells were fixed in 4% paraformaldehyde. After permeabilization and blocking in 5% nonfat dry milk in PBS, the tissue sections or cultured cells were incubated with anti‐IDO antibody (Cell Signaling Technology) or anti‐AhR antibody (Proteintech). Next, tissue sections or cells were incubated with anti‐IgG antibody conjugated with Alexa Fluor 488 (Invitrogen). Rhodamine‐phalloidin (KeyGen) was used for stain actin filaments. 4′,6‐diamidino‐2‐phenylindole (DAPI) (Beyotime) was used to stain the nuclei. Pictures of immunofluorescent staining were visualized with confocal laser microscopy (LSM980, Zeiss).

Anti-AhR antibody (Cat No.: 17840-1-AP) was purchased from Proteintech (Wuhan, China). Anti-NEP antibody (Cat No.: sc-46656) was purchased from Santa Cruz (Dallas, TX, USA). Anti-β-actin antibody (Cat No.: AF5001), HRP-labeled donkey anti-rabbit IgG (Cat No.: A0208) and HRP-labeled donkey anti-mouse IgG (Cat No.: A0216) were purchased from Beyotime Biotechnology (Shanghai, China). Anti-NeuN (Cat No.: ab177487), anti-GFAP (Cat No.: ab7260) and anti-IBA1 (Cat No.: ab178847) antibodies were purchased from Abcam (Cambridge, MA, UK). Alexa Fluor® 488 AffiniPure donkey anti-mouse IgG (H+L) (Code No.: 715-545-150) and Cy™3 AffiniPure donkey anti-rabbit IgG (H+L) (Code No.: 711-165-152) were purchased from Jackson ImmunoResearch (West Grove, PA, USA). The mouse NEP enzyme linked immunosorbent assay (ELISA) kit (Cat No.: CSB-EL014653MO), human Aβ42 ELISA kit (Cat No.: CSB-E10684h) and human Aβ40 ELISA kit (Cat No.: CSB-E08299h) were purchased from Cusabio (Wuhan, China). Diosmin (CAS No.: 520-27-4, Cat No.: HY-N0178), FICZ (CAS No.: 172922-91-7, Cat No.: HY-12451), Indole-3-acetamide (I3C) (CAS No.: 879-37-8, Cat No.: HY-W016784) and Sacubitril (Sac) (CAS No.: 149709-62-6, Cat No.: AHU-377) were purchased from MedChemExpress (Monmouth Junction, NJ, USA). StemRegenin 1 (SR1) (CAS No.: 1227633-49-9, Cat No.: S2858) was purchased from Selleckchem (Shanghai, China).