Western Blotting of Apoptosis Regulators

Cells and tissues were lysed in ice-cold modified RIPA buffer (10 mm Tris-HCl, pH 7.5, 150 mm NaCl, 1 mm EDTA, 1% Igepal), as previously described^{30 (link)}. Protein samples were subjected to electrophoresis on NuPAGE 4–12% polyacrylamide gel (Thermo Fisher Scientific) and then transferred onto a nitrocellulose membrane (BioRad, CA, USA). Equal amounts of protein were Western blotted using the following antibodies: IBtk (#A303-001A; Bethyl Laboratories, Inc., Montgomery, TX, USA), GAPDH (#sc-47724; Santa-Cruz Biotechnology, Dallas, TX, USA), p53 (#sc-393031; Santa-Cruz Biotechnology), Bcl-X_L (#2762, Cell Signaling Technology), c-Myc (#5605, Cell Signaling Technology), Bcl-2 (#7382, Santa-Cruz Biotechnology), Bim (#2933, Cell Signaling Technology), Mcl-1 (#D35A5 Cell Signaling Technology), p19ARF (#sc-32748; Santa-Cruz Biotechnology), Vinculin (V9131, Sigma-Aldrich).

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Vecchio E., Golino G., Pisano A., Albano F., Falcone C., Ceglia S., Iaccino E., Mimmi S., Fiume G., Giurato G., Britti D., Scala G, & Quinto I. (2019). IBTK contributes to B-cell lymphomagenesis in Eμ-myc transgenic mice conferring resistance to apoptosis. Cell Death & Disease, 10(4), 320.

Publication 2019

NuPAGE 4–12% polyacrylamide gel nitrocellulose membrane GAPDH Bcl-XL c-Myc Bcl-2 Mcl-1 p19ARF Vinculin

Antibodies Bcl 2 Buffer C myc Cells Cold Edta Electrophoresis Gapdh Membrane Nacl Nitrocellulose P19arf Polyacrylamide gel Protein Ripa Tissues Tris Vinculin

Corresponding Organization : Magna Graecia University

Other organizations : University of Salerno

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Variable analysis

independent variables

Cells and tissues were lysed in ice-cold modified RIPA buffer (10 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1 mM EDTA, 1% Igepal)

dependent variables

Protein samples were subjected to electrophoresis on NuPAGE 4–12% polyacrylamide gel and then transferred onto a nitrocellulose membrane
Equal amounts of protein were Western blotted using the following antibodies: IBtk, GAPDH, p53, Bcl-XL, c-Myc, Bcl-2, Bim, Mcl-1, p19ARF, Vinculin

control variables

Equal amounts of protein were used for Western blotting

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