Rabbit monoclonal anti tomm20 antibody

For DNA double-strand break analysis, at 18 h after HSV-1 infection, cells were washed with PBS, fixed in 2 to 4% ice-cold paraformaldehyde (Electron Microscopy Sciences) for 10 min, and permeabilized in 90% ice-cold methanol (Sigma) for 30 min. After PBS washing, cells were incubated with 1:100 diluted Alexa Fluor 647 mouse anti-γH2A.X (pS139) antibody (BD Pharmingen) in PBS–5% BSA for 1 h. Mitochondrial quantification was performed by staining EtBr-treated and nontreated THP-1 cells with the rabbit monoclonal anti-TOMM20 antibody (Abcam) diluted 1/40 in PBS–5% BSA for 1 h. After PBS washings, cells were incubated with 1/100-diluted Alexa Fluor 488 goat anti-rabbit in PBS–5% BSA for 45 min. For apoptosis analysis, HeLa and THP-1 cells were collected and washed with PBS, stained with 7-AAD viability staining solution (1/40; eBioscience), then washed with annexin V binding buffer (eBioscience), and stained by anti-annexin V eFluor 450 (1/20; eBioscience). Treatment with 100 μM etoposide in dimethyl sulfoxide was used as positive control. All stained samples were acquired on a MACSQuant analyzer (Miltenyi Biotech), and data were analyzed with FlowJo software (Tree Star, Inc.; version 8.7.1).

OptiPrep™ Density Gradient Medium, MgSO₄, EDTA, HEPES, MES, and KOH were purchased from Sigma (St. Louis, MO, USA). EGTA was purchased from Amresco (USA). Sucrose was purchased from Beijing Biodee Biotechnology (Beijing, China).
The following antibodies were used for immunofluorescence and Western blot analysis: mouse monoclonal anti-proinsulin was purchased from HyTest (Turku, Finland). Guinea pig polyclonal antibody against insulin and rabbit monoclonal anti-TOMM20 antibody were purchased from Abcam (Cambridge, MA). Rabbit polyclonal anti-Vamp4, rabbit polyclonal anti-ATP6V1H, rabbit polyclonal anti-HID-1, rabbit polyclonal anti-CANX, rabbit polyclonal anti-CPE, rabbit polyclonal anti-SYTL4, and rabbit polyclonal anti-STX6 antibodies were purchased from Proteintech (Wuhan, China). Rabbit polyclonal anti-TM9SF3 antibody was purchased from Invitrogen (Carlsbad, CA, USA). Rabbit polyclonal anti-TM9SF3 antibody was purchased from Sigma (St. Louis, MO, USA).