Ab242384

Samples were lysed with Laemmli buffer directly and were run on 10% sodium dodecyl sulfate (SDS)-polyacrylamide gels with a 5% stacking gel. Proteins were then transferred overnight at 20 V to Hybond nitrocellulose membranes (Amersham Biosciences), which were then blocked for 1 hour in 5% w/v skim milk dissolved in TBS (5% milk-TBS) at room temperature, followed by overnight incubation at 4°C with the following primary antibodies diluted in 5% milk-TBS: α-ACE2 (ab108209, 1:500; Abcam), α-TMPRSS2 (ab242384, 1:1000; Abcam), α-actin, (ab8227, 1:2000; Abcam), α-HCMV IE1/2 (11-003, 1:1000; Argene), and α-HSV-1 ICP0 (mouse monoclonal antibody hybridoma supernatant, 11060, 1:5). Blots were washed 3 times in TBS with 0.5% Tween20 (TBS-Tween), and then incubated with secondary antibodies conjugated to horseradish peroxidase. Blots were washed 3 times in TBS-Tween and then incubated with Amersham ECL Western Blotting detection reagent (GE Healthcare Life Sciences) for 5 minutes before being exposed to autoradiography film (Fujifilm).