Western Blot Analysis of HIV Proteins

Protein samples were separated by SDS-polyacrylamide gel electrophoresis and transferred onto PVDF membrane as described elsewhere [31 (link)]. The following antibodies were used: anti-Gag rabbit pAb (Bio Academia), anti-Vpr mouse mAb (Cosmo Bio Co., LTD), anti-FLAG M2 mouse mAb (Sigma-Aldrich), anti-GST rabbit pAb (Medical and Biological Laboratories Co., LTD), anti-GST goat pAb (GE Healthcare), anti-β-actin mouse mAb (Sigma-Aldrich), horseradish-peroxidase (HRP)-conjugated goat anti-mouse IgG (Amersham Biosciences), HRP-conjugated goat anti-rabbit IgG (Amersham Biosciences), and HRP-conjugated donkey anti-goat IgG (Santa Cruz Biotechnology). The intensity of the protein bands was measured using an ImageJ v.1.50 software.

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Chutiwitoonchai N., Siarot L., Takeda E., Shioda T., Ueda M, & Aida Y. (2016). HIV-1 Vpr Abrogates the Effect of TSG101 Overexpression to Support Virus Release. PLoS ONE, 11(9), e0163100.

Publication 2016

anti-FLAG M2 mouse mAb anti-β-actin mouse mAb horseradish-peroxidase (HRP)-conjugated goat anti-mouse IgG HRP-conjugated goat anti-rabbit IgG HRP-conjugated donkey anti-goat IgG

Actin Anti igg Antibodies Biological Donkey Goat Horseradish peroxidase Membrane Mouse Protein Pvdf Rabbit Sds polyacrylamide gel electrophoresis

Corresponding Organization : Osaka University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein band intensity measured using ImageJ v.1.50 software

control variables

None explicitly mentioned

controls

Positive control: None mentioned
Negative control: None mentioned

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