Western Blot Analysis of STAT Proteins

Whole-cell Brij protein lysis and Western-blot analyses were performed as described [46 (link),54 (link)]. Antibodies used for the detection of the respective proteins and their dilutions were: FLAG (M2, SIGMA F-1804; mouse monoclonal; 1:500), p STAT1 (sc-135648, Santa-Cruz Biotechnology; rabbit polyclonal; 1:1000), pSTAT2 (sc-21689-R, Santa-Cruz Biotechnology; rabbit polyclonal; 1:1000), STAT1 (C-terminus, S21120, BD Transduction Laboratories; mouse monoclonal; 1:1000), STAT2 (C-20, sc-476, Santa-Cruz Biotechnology; rabbit polyclonal; 1:1000), pSTAT5 (#9351, Cell Signaling Technology; rabbit polyclonal; 1:1000), STAT5A (L-20, sc-1081, Santa-Cruz Biotechnology; rabbit polyclonal; 1:1000), STAT5A+B (C-17, sc-835, Santa-Cruz Biotechnology; rabbit polyclonal; 1:1000), α-tubulin (DM1A, sc-32293, Santa-Cruz Biotechnology; mouse monoclonal; 1:200), Anti-Rabbit IgG-Peroxidase (SIGMA A-0545; 1:10,000), Anti-Mouse IgG-Peroxidase (SIGMA A-8924; 1:10,000). Apparent molecular weight of detected proteins was as predicted by the antibody manufacturers. Immunoblots shown are representative of at least two independent experiments. Uncropped original blots are available in the S1 File.

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Michl C., Vivarelli F., Weigl J., De Nicola G.R., Canistro D., Paolini M., Iori R, & Rascle A. (2016). The Chemopreventive Phytochemical Moringin Isolated from Moringa oleifera Seeds Inhibits JAK/STAT Signaling. PLoS ONE, 11(6), e0157430.

Publication 2016

pSTAT1 pSTAT2 STAT1 pSTAT5 Anti-Rabbit IgG-Peroxidase Anti-Mouse IgG-Peroxidase

Anti igg Antibodies Antibody Cell Dilutions Immunoblots Mouse Peroxidase Proteins Rabbit Stat1 Stat2 Stat5a Western blot analyses α tubulin

Corresponding Organization : Research Centre for Cereal and Industrial Crops

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Variable analysis

independent variables

Whole-cell Brij protein lysis and Western-blot analyses

dependent variables

Detection of the respective proteins

control variables

Antibody dilutions used for detection of the respective proteins

positive controls

Uncropped original blots available in the S1 File

negative controls

Not explicitly mentioned

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