Rabbit anti giantin antibody

HEK293 (Delecluse et al., 1998 (link)) and HeLa (Kawashima et al., 2013 (link)) cells were maintained in Dulbecco’s modified Eagle medium (DMEM, Gibco BRL) supplemented with 10% fetal bovine serum (FBS). HEK293 cells infected with EBV BRRF2 knockout virus (Watanabe et al., 2015b (link)) were cultured with DMEM containing 150 μg/ml hygromycin B. Akata EBV negative cells (Akata(-)), B95-8, and AGS-EBV cells (Katsumura et al., 2009 (link)) were cultured in RPMI 1640 medium containing 10% FBS.
Rabbit antibodies against BZLF1 and BALF2 were used as reported previously (Murata et al., 2009 (link); Sugimoto et al., 2013 (link)). Mouse anti-FLAG antibody was purchased from Sigma-Aldrich, and rabbit anti-Giantin antibody was from Abcam. Rabbit Anti-tubulin, -Calnexin, and -Rab5 antibodies were purchased from Cell Signaling Technology. Mouse anti-BMRF1 antibody was purchased from Novocastra. Horseradish peroxidase-linked goat antibodies to mouse or rabbit IgG were from Amersham Biosciences.

Tissues were fixed in 4% paraformaldehyde overnight at 4 °C. For the IddU and cell spacing analyses, specimens were embedded in wax by standard methods, sectioned at 8 μm thickness and immunostained according to the manufacturer's instructions with anti‐IddU antibody (BD Biosciences) diluted 1 : 200. For Golgi orientation analyses, specimens were embedded in sucrose–gelatin and thick‐sectioned at 30 μm and immunostained according to the manufacturer's instructions with rabbit‐anti‐giantin antibody (Abcam) at a dilution of 1 : 500. Sections were stained with DAPI nuclear stain before mounting in Mowiol.