PTEN Protein Detection by Western Blot

The SDS-PAGE and Western blot analyses were conducted as previously described [11 (link),13 (link)]. Briefly, after denatured proteins were transferred to a PVDF membrane, the membrane was blocked and probed by rabbit anti-PTEN antibody (Santa Cruz Biotechnology, Santa Cruz, CA). Specific reactions between different antibodies and corresponding proteins were detected by using goat anti-rabbit conjugated with horseradish peroxidase (Sigma, St. Louis, MO) and revealed by a chemiluminescence substrate (Thermo Fisher Scientific, Waltham, MA). The membrane was also probed with anti-Tubulin antibody (Santa Cruz) to normalize the total protein loading. The chemiluminescence signal was digitally recorded and analyzed by the ChemiDoc XRS imaging system (Bio-Rad, Hercules, CA) with Quantity One Program (Version 4.6, Bio-Rad).

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Huang S., Li X, & Zhu H. (2016). MicroRNA-152 Targets Phosphatase and Tensin Homolog to Inhibit Apoptosis and Promote Cell Migration of Nasopharyngeal Carcinoma Cells. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 22, 4330-4337.

Publication 2016

horseradish peroxidase chemiluminescence substrate anti-Tubulin antibody ChemiDoc XRS imaging system Quantity One Program (Version 4.6,

Anti antibody Antibodies Chemiluminescence Goat Horseradish peroxidase Membrane Protein Pten Pvdf Rabbit Sds page Tubulin Western blot analyses

Corresponding Organization :

Other organizations : Zhengzhou Central Hospital, The Sixth People's Hospital of Zhengzhou, Sun Yat-sen University, Sun Yat-sen University Cancer Center

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

PTEN protein expression

control variables

Tubulin protein expression (used to normalize total protein loading)

controls

Positive control: Rabbit anti-PTEN antibody
Negative control: Not explicitly mentioned

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