Western Analysis of Cell Signaling Proteins

Western analysis was carried out as previously described [38 (link)]. Primary antibodies used were p53 1:1000 (NCL-L-p53-DO7, Leica Biosystems Ltd, Newcastle upon Tyne, UK), MYCN 1:500 (sc-53993, Santa Cruz Biotechnology Inc., Dallas, TX, USA), MDM2 1:200 (OP46, Merck), p21^WAF1 1:200 (OP64, Merck), p53 upregulated modulator of apoptosis (PUMA) 1:500 (ab9643, Abcam, Cambridge, UK) and GAPDH 1:500 (sc-25778, Santa Cruz). Experiments were at least n=3.

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Chen L., Rousseau R.F., Middleton S.A., Nichols G.L., Newell D.R., Lunec J, & Tweddle D.A. (2015). Pre-clinical evaluation of the MDM2-p53 antagonist RG7388 alone and in combination with chemotherapy in neuroblastoma. Oncotarget, 6(12), 10207-10221.

Publication 2015

NCL-L-p53-DO7 sc-53993 ab9643 sc-25778

Antibodies Apoptosis Gapdh Mdm2 Mycn

Corresponding Organization :

Other organizations : Newcastle University, La Roche College

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Variable analysis

independent variables

Primary antibodies used were p53 1:1000 (NCL-L-p53-DO7, Leica Biosystems Ltd, Newcastle upon Tyne, UK), MYCN 1:500 (sc-53993, Santa Cruz Biotechnology Inc., Dallas, TX, USA), MDM2 1:200 (OP46, Merck), p21^WAF1 1:200 (OP64, Merck), p53 upregulated modulator of apoptosis (PUMA) 1:500 (ab9643, Abcam, Cambridge, UK) and GAPDH 1:500 (sc-25778, Santa Cruz)

dependent variables

Protein expression levels of p53, MYCN, MDM2, p21^WAF1, PUMA, and GAPDH

control variables

Western analysis was carried out as previously described [38 (link)]

controls

Positive controls: Not specified
Negative controls: Not specified

Annotations

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