Western Blotting of Avian Leukosis Virus

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).

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Xie T., Feng M., Dai M., Mo G., Ruan Z., Wang G., Shi M, & Zhang X. (2019). Cholesterol-25-hydroxylase Is a Chicken ISG That Restricts ALV-J Infection by Producing 25-hydroxycholesterol. Viruses, 11(6), 498.

Publication 2019

rabbit anti-β-actin antibody mouse anti-flag monoclonal antibody IRDye 800-conjugated anti-mouse IgG IRDye 700DX-conjugated anti-rabbit IgG Odyssey FC infrared imaging system

Actin Anti antibody Anti igg Antibody Envelope protein Irdye 800 Monoclonal antibody Mouse Rabbit

Corresponding Organization : South China Agricultural University

Other organizations : Virginia–Maryland College of Veterinary Medicine, University of Maryland, College Park

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Variable analysis

independent variables

ALV-J envelope protein specific mouse anti-monoclonal antibody JE9
Rabbit anti-β-actin antibody
Mouse anti-flag monoclonal antibody

dependent variables

Protein expression levels

control variables

Western blotting protocol described in the referenced publication [26]

controls

Positive control: Not specified
Negative control: Not specified

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