Anti flag ma1 91878

Cells were lysed in a radioimmunoprecipitation assay (RIPA; solarbio) buffer containing 1mM phenylmethylsulfonyl fluoride and protease inhibitors (Sigma-Aldrich), and immunoblotting was performed as previously described [31 (link)]. The antibodies used in this study were as follows: anti-GAPDH (AB0036; Abways), anti-GBP1 (bs-13302R; Bioss), anti-HA (66006-2-Ig; Proteintech), anti-Flag (MA1-91878; Thermo Fisher), anti-Flag (701,629; Invitrogen), horseradish peroxidase (HRP)-conjugated anti-mouse IgG and anti-rabbit IgG (A16066 and 31 466; Invitrogen), anti-mouse IgG (H&L)-Alexa Fluor 488 (RS3208; Immunology), anti-mouse IgG (H&L)-Alexa Fluor 647 (RS3808; Immunology), anti-rabbit IgG (H&L)-Alexa Fluor 647 (RS3811; Immunology), anti-rabbit IgG (H&L)-Alexa Fluor 594 (RS3611; Immunology). Anti-gC and anti-EP0 of PRV polyclonal antibodies were produced in our lab.

The methods for protein measurement in cell lysates and Western blotting were performed as described previously (Chen et al., 2018 (link)). The antibodies for immunoblots used in this study are follows: anti-HBc (B0586, Dako, Denmark), anti-flag (MA-1-91878, Thermo, USA), anti-ODC1(sc-398116, Santa Cruz, USA), anti-SRM (bs-17653R, Bioss, China), anti-elF5A (ET1610-49, Hangzhou Hua An Biotechnology, China), anti-HBs (NB100-62652, Novus, USA), anti-GAPDH (100242-MM05, Sino Biological, China). Quantifications of the immunoblot band intensities were analyzed by the Image J software.

For western blotting of cell lysate, the cells were lysed in the same lysis buffer as that used in immunoprecipitation. For the analysis of the immunoprecipitation, the protein-bound beads were heated in Laemmli sample buffer at 95 °C for 5 min. The sample was analyzed by western blotting, as previously described [34 (link)]. The antibodies to glutaminase (GLS) (ab93434), glutamate dehydrogenase (GDH) (ab55061) and CAD (ab99312) were available from Abcam (Cambridge, UK). The antibody to actin (clone AC-40, A3853) was purchased from Sigma-Aldrich (St. Louis, Missouri, USA). The horseradish peroxidase (HRP)-conjugated secondary antibodies to rabbit (sc-2004) and mouse (sc-2005) IgG were available from Santa Cruz Biotechnology Inc. (Dallas, Texas, USA). The HRP-conjugated anti-c-Myc (R951-25) and anti-FLAG (MA1-91878) antibodies were available from Thermo Fisher Scientific. Rabbit polyclonal anti-EV71-VP1 antibody (PAB7631-D01P) was purchased from Abnova (Neihu District, Taipei City, Taiwan).