Western Blot Protein Analysis Protocol

Immunoblots were prepared with 30–50 µg of protein lysate/sample, as previously described (Tan et al., 2016 (link)). Antibodies used for immunoblotting included a pan-Akt antibody (Cell Signaling Technology, Cat# 9272, RRID: AB_329827); anti-P-Akt (Ser473) antibody (Cell Signaling Technology, Cat# 4060, RRID: AB_2315049); rabbit anti-c-Myc XP monoclonal antibody, unconjugated, clone D84C12 (Cell Signaling Technology, Cat# 5605S, RRID: AB_1903938); p53 mouse 1C12 monoclonal antibody (Cell Signaling Technology, Cat# 2524, RRID: AB_331743); rabbit anti-PTEN monoclonal antibody, unconjugated, clone 138G6 (Cell Signaling Technology, Cat# 9559, RRID: AB_390810); anti-P-S6 ribosomal protein (Ser235/236) (91B2) rabbit monoclonal antibody (Cell Signaling Technology, Cat# 4857, RRID: AB_2181035); mouse anti-Gapdh monoclonal antibody, unconjugated, clone 6c5 (Santa Cruz Biotechnology, Cat# sc-32233, RRID: AB_627679); BAP1 antibody (Bethyl Cat# A302-243A, RRID: AB_1731059); NF2/merlin (H-260) antibody (Santa Cruz Biotechnology, Cat# sc-28247, RRID: AB_2149822).

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Sementino E., Menges C.W., Kadariya Y., Peri S., Xu J., Liu Z., Wilkes R.G., Cai K.Q., Rauscher FJ I.I.I., Klein-Szanto A.J, & Testa J.R. (2018). Inactivation of Tp53 and Pten drives rapid development of pleural and peritoneal malignant mesotheliomas. Journal of cellular physiology, 233(11), 8952-8961.

Publication 2018

pan-Akt antibody anti-P-Akt (Ser473) antibody

Anti antibody Anti c antibody Antibodies Antibody Clone Gapdh Merlin Monoclonal antibody Mouse Protein Pten Rabbit Ribosomal protein

Corresponding Organization : Fox Chase Cancer Center

Other organizations : The Wistar Institute

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Variable analysis

independent variables

Protein lysate concentration (30-50 µg)

dependent variables

Levels of pan-Akt
Levels of phosphorylated Akt (Ser473)
Levels of c-Myc
Levels of p53
Levels of PTEN
Levels of phosphorylated S6 ribosomal protein (Ser235/236)
Levels of GAPDH
Levels of BAP1
Levels of NF2/merlin

control variables

Sample preparation and immunoblotting procedures as previously described (Tan et al., 2016)

controls

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

Annotations

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