Alexa fluor 488 goat anti rabbit igg

RIF; INH; trans-2-pentenal; fluoresceinamine isomer 1; poly(d,l-lactide-coglycolide) acid (PLGA) terminated, lactide:glycolide 50:50; and Float-A-Lyser G2 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Pooled human serum was purchased from Innovative Biologics (Herndon, VA, USA). Macrophage colony stimulating factor (MCSF) was prepared from culture fluids recovered from 5/9m α3–18 cells [CRL-10154; American Type Culture Collection (ATCC), Manassas, VA, USA] cultured in ATCC complete growth medium (35 ). Rabbit anti-human antibodies to Rab 5, 7, 11, and 14 and Alexa Fluor 488 goat anti-rabbit IgG were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Protein A/G mix magnetic beads were purchased from Millipore (Billerica, MA, USA).

Gallium (III) meso tetraphenylporphyrine chloride was purchased from Frontier Scientific (Logan, Utah, USA). Fluoresceinamine isomer 1; and Float-A-Lyser G2 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Pooled human serum was purchased from Innovative Biologics (Herndon, VA, USA). Macrophage colony stimulating factor (MCSF) was prepared from 5/9m alpha3-18 cells (ATCC®, CRL-10154™) cultured in ATCC complete growth medium30 (link). Rabbit anti-human antibodies to Rab 5, 7, 11 and 14, and Alexa Fluor 488 goat anti-rabbit IgG were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

5×10⁴ cells, with or without VX680 for 48 h, were seeded into millicell ez slides (Millipore, MA, USA) and fixed with 4% paraformaldehyde (PFA) for 30 min. Slides were rinsed three times with PBS, blocked with 5% BSA in PBS containing 0.05% Triton for 1 h at room temperature and incubated overnight with primary antibodies at 4°C. Slides were then rinsed three times with PBS and incubated with secondary antibodies for 1 h at room temperature in the dark. Nuclei were visualized with DAPI (1:1000, Beyotime) in PBS for 5 min in the dark. Slides were rinsed three times in PBS and analyzed by fluorescent microscopy (10x). Primary antibodies included anti-p-AURKA (1:100, Anti-rabbit IgG, Cell Signaling Technology), anti-P130 (1:100, Anti-rabbit IgG, Santa Cruz) and anti-P107 (1:100, Anti-rabbit IgG, Santa Cruz). Secondary antibodies included Alexa Fluor® 488 goat Anti-rabbit IgG and Alexa Fluor® 555 goat Anti-rabbit IgG (1:1000, Santa Cruz).