Protein Expression Profiling in A673/TR/shEF1 Cells
Protein from A673/TR/shEF1 cells was extracted at d0, d7, d11, d14, and d17 with RIPA and anti-protease cocktail (Roche). Western blots were performed following routine protocols and specific band detection was achieved by the use of rabbit monoclonal anti-FLI1 antibody (1:1000, ab133485; Abcam)38 (link), rabbit polyclonal anti-MYBL2 antibody (1:500, sc-725; Santa Cruz)39 (link), and mouse monoclonal anti-ß-actin (1:10,000, A-5316; Sigma-Aldrich). Anti-rabbit IgG horseradish peroxidase-coupled antibody (1:3000, Amersham Bioscience) and anti-mouse IgG horseradish peroxidase coupled antibody (1:3,000; Amersham Bioscience) was used as secondary antibody. Proteins were visualized using chemiluminescence (Pierce ECL Western blot chemiluminescent substrate; Thermo Fisher Scientific).
Other organizations :
Ludwig-Maximilians-Universität München, Institut Curie, Inserm, Université Paris Sciences et Lettres, Ben-Gurion University of the Negev, German Cancer Research Center, Heidelberg University, University Hospital Münster, Essen University Hospital, University of California, San Diego, Hospital for Sick Children, LMU Klinikum
Protein extraction protocol (RIPA and anti-protease cocktail)
Western blotting protocol
Antibodies used for detection (anti-FLI1, anti-MYBL2, anti-β-actin)
Secondary antibodies used (anti-rabbit and anti-mouse IgG)
controls
Positive control: β-actin for normalizing protein expression levels
Negative control: Not explicitly mentioned
Annotations
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