Protein Expression Analysis by SDS-PAGE and Immunoblot

TCEs were separated on 10% SDS-PAGE gels, transferred to nitrocellulose membranes, incubated first with the primary antibodies against proteins or protein phosphor-isoforms of interest (Supplemental Table S1) and then with IRDye 680RD secondary antibodies (cat. no. 92668070, LI-COR, Lincoln, NE). Immunoblots were detected using the LI-COR’s Odyssey CLx imaging system. Membranes were stripped and re-incubated with antibodies against β-actin, which served as a loading control.

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Urasaki Y., Fiscus R.R, & Le T.T. (2018). Detection of the Cell Cycle-Regulated Negative Feedback Phosphorylation of Mitogen-Activated Protein Kinases in Breast Carcinoma using Nanofluidic Proteomics. Scientific Reports, 8, 9991.

Publication 2018

Odyssey CLx

Actin Antibodies Gels Immunoblots Membranes Nitrocellulose Phosphor Protein isoforms Proteins Sds page Tces

Corresponding Organization :

Other organizations : Roseman University of Health Sciences

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Variable analysis

independent variables

Primary antibodies against proteins or protein phosphor-isoforms of interest (Supplemental Table S1)

dependent variables

Protein or protein phosphor-isoform levels

control variables

β-actin (used as a loading control)

controls

Positive control: None specified
Negative control: None specified

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