Western Blot Analysis of Transfected Cells

Cells that were transfected with individual reporters were lysed with Lysis Buffer (50 mM Tris pH 7.8, 150 mM NaCl, 1% NP-40). Western blot analysis was performed, as described previously^{64 (link)}. The following primary antibodies were used in western analysis at the given dilution: GFP, 1:2000 (Clontech, 632381); β-actin-HRP, 1:2000, (Cell Signaling, 12262); Anti-mouse IgG HRP, 1:10,000 (Cell Signaling, 7076S).

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Cottrell K.A., Chaudhari H.G., Cohen B.A, & Djuranovic S. (2018). PTRE-seq reveals mechanism and interactions of RNA binding proteins and miRNAs. Nature Communications, 9, 301.

Publication 2018

β-actin-HRP Anti-mouse IgG HRP

Actin Anti igg Antibodies Buffer Cells Dilution Mouse Nacl Np 40 Tris Western blot analysis

Corresponding Organization : Washington University in St. Louis

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Variable analysis

independent variables

Transfection with individual reporters

dependent variables

Protein expression levels of GFP, β-actin

control variables

Lysis Buffer (50 mM Tris pH 7.8, 150 mM NaCl, 1% NP-40)
Western blot analysis as described previously in reference 64

controls

Positive control: None specified
Negative control: None specified

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