Rabbit anti β actin antibody

Chicken MDM were infected with a 10⁵ TCID₅₀/mL of ALV-J strain SCAU-HN06. DNA, RNA and total proteins were extracted from the ALV-J infected MDM at 3, 6, 12, 24 and 36 h post-infection (hpi). RT-PCR was employed to detect the ALV-J replication using specific PCR primers H5/H7 [12 (link)]. Western blotting was performed with ALV-J envelope protein specific mouse antibody JE9 (kindly provided by Dr Aijian Qin, Yangzhou University, Yangzhou, China) and rabbit anti-β-actin antibody (Bioworld, Louis Park, USA) according to the method described previously [13 (link)]. IRDye 700DX-conjugated anti-rabbit IgG and IRDye 800-conjugated anti-mouse IgG (Rockland Immunochemicals, Limerick, PA, USA) was used as the secondary antibody. Membranes were visualized and analyzed with an Odyssey infrared imaging system (LI-COR Biosciences, Lincoln, NE, USA). ALV-J provirus was detected by PCR with primers H5/H7 using DNA template.

Western blotting was performed with ALV-J envelope protein specific mouse anti-monoclonal antibody JE9 (kindly provided by Prof. Aijian Qin, Yangzhou University, Yangzhou, China), rabbit anti-β-actin antibody (Bioworld, Louis Park, MN, USA) and mouse anti-flag monoclonal antibody (Proteintech, Rosemont, CA, USA) according to our previously described method [26 (link)]. IRDye 800-conjugated anti-mouse IgG and IRDye 700DX-conjugated anti-rabbit IgG (Rockland Immunochemicals, Limerick, PA, PA, USA) were used as the secondary antibody. Results were visualized and analyzed with an Odyssey FC infrared imaging system (LICOR Biosciences, Lincoln, NE, USA).