Immunoblot Analysis of Melanoma Cell Proteins

Whole-cell lysates of melanoma cells and nuclear-enriched cell lysates were prepared and immunoblot analysis was performed as described previously^{22 (link)}. Fifteen μg protein of whole-cell lysate or 5 μg of nuclear-enriched lysates were used for the sodium dodecyl sulphate–polyacrylamide gel electrophoresis. The following primary antibodies were used: anti-Lamin-B (C-20, Santa Cruz Biotechnology, 1:250), anti-p53 (DO-I, Santa Cruz Technology), anti-Actin (D6A8, CST, 1:1000), anti-p73 (GC-15, BD Biosciences, 1:250; H-79, Santa Cruz, 1:250; 5B429, Novus Biologicals, 1:500), anti- pH2AX (JBW301, Abcam, 1:500), anti-Mdm2 (Abcam, 1: 100), anti-p21 (CST, 1:2000), and anti-HA (Santa Cruz, 1:250).

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Makino E., Gutmann V., Kosnopfel C., Niessner H., Forschner A., Garbe C., Sinnberg T, & Schittek B. (2018). Melanoma cells resistant towards MAPK inhibitors exhibit reduced TAp73 expression mediating enhanced sensitivity to platinum-based drugs. Cell Death & Disease, 9(9), 930.

Publication 2018

anti-Lamin-B (C-20, anti-p53 (DO-I, anti-Mdm2 anti-HA

Actin Antibodies Biologicals Cell Immunoblot analysis Lamin b Mdm2 Melanoma Novus Protein Sodium dodecyl sulphate polyacrylamide gel electrophoresis

Corresponding Organization :

Other organizations : University of Tübingen

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Variable analysis

independent variables

Whole-cell lysates of melanoma cells
Nuclear-enriched cell lysates

dependent variables

Protein expression levels of Lamin-B
Protein expression levels of p53
Protein expression levels of Actin
Protein expression levels of p73
Protein expression levels of pH2AX
Protein expression levels of Mdm2
Protein expression levels of p21
Protein expression levels of HA

control variables

Protein amount used for sodium dodecyl sulphate–polyacrylamide gel electrophoresis (15 μg for whole-cell lysates, 5 μg for nuclear-enriched lysates)

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