Alexa fluor 488 goat anti rabbit a0423

hMOF-overexpressed OVCAR3 cells were seeded into plates containing glass coverslips and cultured for 24 h. The cell climbing slice was fixed with 4% paraformaldehyde for 2 h, permeabilized in 0.5% PBST (PBS containing 0.5% Triton X-100) for 10 min and blocked with 5% goat serum for 60 min. Then the cell slides were incubated with primary antibodies of hMOF (#ab200660, 1:1000, Abcam) and MDM2 (#66511-1-Ig, 1:500, Proteintech) at 4 °C overnight. After incubation with secondary antibodies (Alexa fluor 488 goat anti-rabbit, A0423, Beyotime, 1:1000; Alexa fluor 555 donkey anti-mouse, A0460, Beyotime, 1:1000) for 2 h at room temperature, the nuclei were counterstained with DAPI (1 μg/mL) for 5 min. The immunofluorescence images were captured by a Nikon NiE-A1 plus or Nikon C2+ confocal microscope, and analyzed using Nikon-elements AR software.

Antibodies: AKT (4691), P38 (8690), ERK1/2 (4695), p-AKT (4060), p-P38 (4511), p-ERK1/2 (4370), IκBα (4814), p-IκBα (2859), p-IKKα/β (2697), NF-κB (8242), Rab5a (46449), horse anti-mouse IgG (H&L) (7076), and goat anti-rabbit IgG (H&L) (7074) were purchased from Cell Signaling Technology (Danvers, MA, United States). IKKβ (ab124957), PBR (ab109497) was from Abcam (CA, United States). Alexa Fluor 488 goat anti-rabbit (A0423) and Alexa Fluor 647 goat anti-mouse (A0473) secondary antibodies were obtained from Beyotime Biotechnology (Shanghai, China). TLR4 (66350-1-Ig), GAPDH (60004-1-Ig), and β-Tubulin (10068-1-AP) were purchased from Protein-Tech (Wuhan, Hubei, China). Lipopolysaccharides(L4391) was obtained from Sigma (Louis, MO, United States), Remimazolam Tosilate for Injection (Remimazolam) were obtained from Hengrui (Jiangsu, China).