Anti histone h3 tri methyl k9 antibody

Protein samples were mixed with Laemmli buffer (BioRad) and β-mercaptoethanol (Sigma). Each sample was analyzed in triplicate. Next, samples were denatured, loaded on a gel (0.1 µg protein per sample), and separated by electrophoresis (60 min, 125 V). Then proteins were blotted onto a polyvinylidene difluoride (PVDF) membrane using a Trans-Blot Turbo Transfer System (BioRad). Membranes were blocked with 3% BSA for 1 h at RT and incubated with primary antibodies (anti-histone H3 (tri methyl K9) antibody (Abcam), anti-histone H4 (tri methyl K20) antibody (Abcam), anti-histone H4 (acetyl K8) antibody (Abcam), anti-histone H3 (tri methyl K4) antibody (Abcam), anti-histone H3 (tri methyl K27) antibody (Abcam), and anti-histone H3 (acetyl K9) antibody (Abcam)) in TBST for 16 h at 4 °C. The membranes were then washed three times for 10 min with TBST, and were incubated with secondary antibody (anti-rabbit) for 1 h at RT and washed. The chemiluminescence reaction was performed with Clarity Western ECL Substrate (BioRad). Signals were visualized with ChemiDoc Imaging System (BioRad), and signal intensities were measured using ImageLab software (BioRad).

Primary Antibodies: anti-CENPA (MBL international 1:500, Code # D115-3), anti-CENPB primary antibody (Santa Cruz Biotechnology, CENPB Antibody (C-10) 1:50: sc-376392, pSer139-γ-H2AX antibody (Cell Signaling 1:400, Cat # 9718), BANF1/BAF antibody (abcam,1:100, EPR7668), MHCII DRB1 antibody (Abclonal, 1:100, Cat # A7685), Lamin B1 antibody (Proteintech, 1:1000, Cat # 66095-1-lg), cGAS antibody (Abclonal, 1:100, Cat# A8335), p(Ser396)-IRF antibody (Cell Signaling, 1:100, Cat # 29047), p(Ser536)-p65 antibody (Abclonal, 1:100, Cat # AP0123), MHCII DRB5 antibody (Abclonal, 1:100, Cat # A12726), anti-GAPDH antibody (Abcam, ab 9484), anti-Histone H3 (tri methyl K9) antibody (Abcam, ab8898), anti-CENPA antibody [3-19] (Abcam, ab13939).
Secondary Antibodies: Alexa Flour 594 rabbit anti-mouse IgG 1:1000, Thermo Fischer Scientific, Cat No A27027, Alexa Flour 488 rabbit anti-mouse IgG 1:1000, Thermo Fischer Scientific, Cat No A27023. Alexa fluor 647 Goat anti-rabbit IgG, ThermoFisher, 1:1000, Cat # A32733TR

Dulbecco’s modified Eagle’s medium (DMEM) and penicillin/streptomycin (PenStrep) were purchased from Gibco–Life Technologies (Carlsbad, CA, USA). Fetal bovine serum (FBS) was purchased from MP Biomedicals (Irvine, CA, USA). Bovine serum albumin (BSA), phosphate buffered saline (PBS), and 40 nm silver nanoparticles (SNPs) were purchased from Sigma-Aldrich (St Louis, MO, USA). The 40 nm gold nanoparticles (GNPs) were purchased from BBI Solutions (Crumlin, UK). Anti-Histone H3 (tri methyl K9) antibody and Anti-Histone H3 (di methyl K4) antibody were purchased from Abcam (Cambridge, MA, USA). Anti-Histone H3 (tri methyl K27) antibody was purchased from Millipore (MA, USA) and TGF-β1 polyclonal antibody from Biorbyt (Cambridge, UK). Goat anti-Rabbit IgG (H + L) cross-adsorbed secondary antibody, Alexa Fluor 488 was purchased from Invitrogen (MA, USA).