The primary antibodies used for immunocytochemistry were as follows: anti-Irgb10 rabbit polyclonal antiserum (64 (link)) at 1:1,000, anti-Gbp2 rabbit polyclonal (54 (link), 64 (link)) at 1:500, anti-LAMP1 mouse monoclonal (Santa Cruz Biotechnology) at 1:250, anti-LAMP1 rabbit polyclonal (Gbiosciences) at 1:250, anti-LAMP2 rabbit polyclonal (Invitrogen) at 1:250, and anti-LC3 rabbit polyclonal (MBL International) at 1:250. Protein samples from whole-cell lysates were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. The blots were probed with primary antibodies specific for the following: anti-Gbp2 rabbit polyclonal at 1:1,000, anti-Irgb10 rabbit polyclonal antiserum at 1:1,000, anti-hGBP1 rat monoclonal at 1:1,000 (Santa Cruz Biotechnology), and anti-β-actin mouse monoclonal (Sigma) at 1:5000. Binding of secondary horseradish peroxidase (HRP)-labeled goat anti-rabbit IgG at 1:2,000 or goat anti-rat IgG at 1:2,000 or goat anti-mouse IgG antibodies at 1:5,000 (GeNei) was analyzed using Clarity Western ECL (Bio-Rad) or Westar Supernova chemiluminescent substrate (Cyanagen).
Haldar A.K., Nigam U., Yamamoto M., Coers J, & Goyal N. (2020). Guanylate Binding Proteins Restrict Leishmania donovani Growth in Nonphagocytic Cells Independent of Parasitophorous Vacuolar Targeting. mBio, 11(4), e01464-20.
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