Protocol detail

Western Blot Quantification Protocol

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Western blotting was carried out largely as described previously (Song et al., 2019 (link)), using the following primary antibodies: rabbit anti-TOX4 (Sigma, HPA017880, 1:1000; and Abcam, ab66651, 1:1000), mouse anti-ACTIN (Abcam, ab3280, 1:5000) and rabbit anti-GAPDH (Sigma, G9545, 1:1000) antibodies. Secondary antibodies were: HRP-conjugated goat anti-mouse-IgG antibody (Bio-Rad, 1706516, 1:5000) or goat anti-rabbit-IgG antibody (Bio-Rad, 1706515, 1:5000) for 30 min at room temperature. Data were analyzed with ImageJ.

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Vanheer L., Song J., De Geest N., Janiszewski A., Talon I., Provenzano C., Oh T., Chappell J, & Pasque V. (2019). Tox4 modulates cell fate reprogramming. Journal of Cell Science, 132(20), jcs232223.

Publication 2019

ab3280 G9545 HRP-conjugated goat anti-mouse-IgG antibody goat anti-rabbit-IgG antibody

Actin Anti igg Antibodies Antibody Gapdh Goat Mouse Rabbit

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Variable analysis

independent variables

Primary antibodies used: rabbit anti-TOX4 (Sigma, HPA017880, 1:1000; and Abcam, ab66651, 1:1000), mouse anti-ACTIN (Abcam, ab3280, 1:5000) and rabbit anti-GAPDH (Sigma, G9545, 1:1000)

dependent variables

Protein expression levels of TOX4, ACTIN, and GAPDH as measured by Western blotting

control variables

Western blotting procedure largely carried out as described previously (Song et al., 2019)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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