Protein Immunodetection by Western Blot

Proteins were separated by SDS-PAGE and transferred onto PVDF membranes. Antibodies used against PTEN were: (138G6, Cell Signaling), GFP45 (link), NEDD4-1 (ab14592, Abcam), p70S6K and phospho-p70S6k duet (8209, Cell Signaling), AKT (9272, Cell Signaling), phospho-AKT (4060, Cell Signaling) and actin (C-11, Santa Cruz Biotechnology). Immunocomplexes were visualized by fluorescently labeled secondary antibodies and detected using a PharosFX Plus molecular imager (Bio-Rad).

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Nguyen H.N., Yang J.M., Miyamoto T., Itoh K., Rho E., Zhang Q., Inoue T., Devreotes P.N., Sesaki H, & Iijima M. (2015). Opening the conformation is a master switch for the dual localization and phosphatase activity of PTEN. Scientific Reports, 5, 12600.

Publication 2015

ab14592 phospho-AKT PharosFX Plus

Actin Antibodies Membranes P70s6k Proteins Pten Pvdf Sds page

Corresponding Organization :

Other organizations : Johns Hopkins University, Johns Hopkins Medicine

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Variable analysis

independent variables

Antibodies used against PTEN, GFP45, NEDD4-1, p70S6K and phospho-p70S6k, AKT, phospho-AKT, and actin

dependent variables

Protein expression levels of PTEN, GFP45, NEDD4-1, p70S6K, phospho-p70S6k, AKT, phospho-AKT, and actin

control variables

SDS-PAGE separation of proteins
Transfer of proteins onto PVDF membranes
Fluorescently labeled secondary antibodies
Detection using PharosFX Plus molecular imager (Bio-Rad)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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