Western Blot Analysis of Cyclic Nucleotide Phosphodiesterases

Lysates were run on either denaturing NuPAGE Novex 4-12% Bis-Tris or NativePAGE Novex 4-16% Bis-Tris gels, as per manufacturer’s instruction (Life Technologies). Blots were probed overnight at 4°C with antibodies recognizing PDE11A (1:500, PD11A-112AP FabGennix—Frisco, TX [4 (link)-6 (link)]), PDE2A (1:1000, PD2A-101AP FabGennix [38 (link)] ), PDE10A (1:2500, Enzo Life Science; Farmingdale, NY [39 (link)]), actin (1:10,000, A2066 Sigma; St. Louis, MO), IL-6 (1:200, MAB406 R&D Systems; Minneapolis, MN [40 ]), GFP (1:2000, sc-8334 Santa Cruz; Dallas, TX), or RFP (1:10,000, A00682 Genscript). Blots were then processed as previously published [4 (link), 6 (link), 41 (link)].

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Pathak G., Agostino M.J., Bishara K., Capell W.R., Fisher J.L., Hegde S., Ibrahim B.A., Pilarzyk K., Sabin C., Tuczkewycz T., Wilson S, & Kelly M.P. (2016). PDE11A negatively regulates lithium responsivity. Molecular psychiatry, 22(12), 1714-1724.

Publication 2016

NuPAGE Novex 4-12% Bis-Tris NativePAGE Novex 4-16% Bis-Tris PDE10A sc-8334 A00682

Actin Antibodies Bis tris Gels Pde2a

Corresponding Organization :

Other organizations : University of South Carolina, Pfizer (United States)

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Variable analysis

independent variables

Denaturing NuPAGE Novex 4-12% Bis-Tris gel
NativePAGE Novex 4-16% Bis-Tris gel

dependent variables

Levels of PDE11A
Levels of PDE2A
Levels of PDE10A
Levels of actin
Levels of IL-6
Levels of GFP
Levels of RFP

control variables

Manufacturer's instructions (Life Technologies)

positive controls

Antibodies recognizing PDE11A, PDE2A, PDE10A, actin, IL-6, GFP, and RFP

negative controls

Not explicitly mentioned

Annotations

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