Western Blot Analysis of SREBP

Colon cancer cells were harvested and detergent-solubilized cell lysates were obtained as described previously^{41 (link)–44 (link)}. Equal amounts of cell lysates were resolved by SDS-PAGE and subjected to western blot analysis. The SREBP1 antibody was from BD Biosciences (CA, USA), whereas the SREBP2 antibody was from R&D Systems (MN, USA). Note that the SREBP antibodies recognize both the precursor and mature active forms of the respective proteins. The γ-tubulin antibody was from Sigma-Aldrich.

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Wen Y.A., Xiong X., Zaytseva Y.Y., Napier D.L., Vallee E., Li A.T., Wang C., Weiss H.L., Evers B.M, & Gao T. (2018). Downregulation of SREBP inhibits tumor growth and initiation by altering cellular metabolism in colon cancer. Cell Death & Disease, 9(3), 265.

Publication 2018

SREBP1 antibody SREBP2 antibody γ-tubulin antibody

Antibodies Antibody Cell Colon cancer Detergent Proteins Sds page Srebp1 Tubulin Western blot analysis

Corresponding Organization :

Other organizations : Markey Cancer Center, University of Kentucky

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Protein expression levels of SREBP1 and SREBP2

control variables

Equal amounts of cell lysates used for SDS-PAGE and western blot analysis
γ-tubulin used as a loading control

controls

Positive control: None specified
Negative control: None specified

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