Western Blot Analysis of Cell Signaling

Western blot was performed as previously described[23 (link)]. Antibodies used were against c-Myc (BD Pharmingen), TRIM3 (GeneTex), β-actin, serving as a loading control (Santa Cruz Biotechnology and GeneTex), p21^cip, p27^kip1, Cyclin D2, Notch, Numb, Hes1, and MS1 (Musashi)(all Cell Signaling). Goat-anti-mouse IgG or goat-anti-rabbit IgG conjugated to horseradish peroxidase (Bio-Rad) were used as secondary antibodies. Enhanced chemiluminescence (ECL) was used for detection (Thermo Scientific).

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Chen G., Kong J., Tucker-Burden C., Anand M., Rong Y., Rahman F., Moreno C.S., Van Meir E.G., Hadjipanayis C.G, & Brat D.J. (2014). Human Brat ortholog TRIM3 is a tumor suppressor that regulates asymmetric cell division in glioblastoma. Cancer research, 74(16), 4536-4548.

Publication 2014

c-Myc TRIM3 β-actin Goat-anti-mouse IgG goat-anti-rabbit IgG conjugated to horseradish peroxidase Enhanced chemiluminescence (ECL)

Actin Anti igg Antibodies Chemiluminescence Cyclin d2 Goat Horseradish peroxidase Mouse P27kip1 Rabbit Western blot

Corresponding Organization : Emory University

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Variable analysis

independent variables

Western blot protocol previously described[23]
Antibodies used against c-Myc, TRIM3, β-actin, p21cip, p27kip1, Cyclin D2, Notch, Numb, Hes1, and MS1 (Musashi)

dependent variables

Protein expression levels of c-Myc, TRIM3, β-actin, p21cip, p27kip1, Cyclin D2, Notch, Numb, Hes1, and MS1 (Musashi)

control variables

β-actin, serving as a loading control

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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