Western Blotting Antibody Validation

Methods for Western blotting was described elsewhere^{16 (link)}. Polyclonal rabbit antibody for Fli-1 were obtained from Abcam (Abcam, Cambridge, UK); ERK and phospho-ERK from Cell Signalling Technology (CST, Danvers, MA01923), β-actin, β-Tubulin, BCL-2 from Proto-Technology (Protein-Tech, Bucuresti, Romania) and goat-anti-mouse, and goat anti-rabbit HRP-conjugated (Promega). Antibody dilution was according to the manufacturer instructions. Imaging of proteins was performed using Oddessy system (Li-Cor biosciences, Lincoln, USA). In experiments described in Fig. 7a, we used ECL detection system for western blotting, as described^{9 (link)}.

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Liu T., Xia L., Yao Y., Yan C., Fan Y., Gajendran B., Yang J., Li Y.J., Chen J., Filmus J., Spaner D.E., Zacksenhaus E., Hao X, & Ben-David Y. (2019). Identification of diterpenoid compounds that interfere with Fli-1 DNA binding to suppress leukemogenesis. Cell Death & Disease, 10(2), 117.

Publication 2019

Oddessy system

Actin Antibody Bcl 2 Dilution Goat Mouse Promega Proteins Rabbit Tubulin

Corresponding Organization :

Other organizations : Guiyang Medical University, Kunming Institute of Botany, Chinese Academy of Sciences, First Bethune Hospital of Jilin University, Sunnybrook Hospital, University of Toronto, Key Laboratory of Chemistry for Natural Products of Guizhou Province and Chinese Academy of Sciences

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Variable analysis

independent variables

Antibody type (Fli-1, ERK, phospho-ERK, β-actin, β-Tubulin, BCL-2)

dependent variables

Protein expression levels

control variables

Antibody dilution (according to manufacturer instructions)
Imaging system (Odyssey system)

controls

Positive control: ECL detection system for Western blotting used in experiments described in Fig. 7a
Negative control: Not explicitly mentioned

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