Hrp conjugated goat anti mouse immunoglobulin

The following primary antibodies were diluted in TBST plus 5% nonfat dry milk for western blot: rabbit anti-mGBP-2 (1851 [21 (link),22 (link)]; 1:20,000), rabbit anti-actin (A2066; 1:3000) (Sigma), rabbit anti-α-tubulin (GTX102078; 1:10,000) (GeneTex, Irvine, CA, USA), mouse anti-GAPDH (60004; 1:20,000) (Proteintech, Rosemont, IL, USA). The above antibodies were incubated with membrane for 1 h at RT. The rabbit anti-RhoA (2117T; 1:4000) (Cell Signaling, Danvers, MA, USA), mouse anti-Rac1 (610650; 1:4000) (BD Biosciences, San Jose, CA, USA), mouse anti-CDC42 (610928; 1:250) (BD Biosciences), rabbit anti-Akt (9272; 1:3000) (Cell Signaling), and rabbit monoclonal anti-phosphoAkt (4060 (ser 473); 1:4000) (Cell Signaling) were incubated with membrane overnight at 4 °C. Hrp-conjugated goat anti-mouse immunoglobulin (1:8000) or goat anti-rabbit immunoglobulin (1:6000) (Jackson ImmunoResearch, West Grove, PA, USA) were diluted in TBST containing 5% w/v nonfat dry milk and incubated with the membranes for 1 h at RT.