Western Blot and Immunostaining Analysis

Western blot were carried out as previously described [7 (link)]. Immunostaining was carried out using a rabbit monoclonal caspase3, caspase9, actin and PhosphoAKT antibody (1/1000, Cells signaling) and a secondary polyclonal mouse anti-rabbit antibody HRP conjugated (1/2000, cell signalling). Blots were developed using HRP and chemiluminescent peroxidase substrate (#CPS1120, Sigma). Data were collected using Geliance CCD camera (Perkin Elmer), and analyzed using ImageJ software (NIH).

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Pasquier J., Vidal F., Hoarau-Véchot J., Bonneau C., Daraï E., Touboul C, & Rafii A. (2018). Surgical peritoneal stress creates a pro-metastatic niche promoting resistance to apoptosis via IL-8. Journal of Translational Medicine, 16, 271.

Publication 2018

Geliance CCD camera

Actin Anti antibody Antibody Caspase3 Cells Mouse Peroxidase Rabbit Western blot

Corresponding Organization : Hôpital Foch

Other organizations : Inserm, Hôpital Tenon, Sorbonne Université, Assistance Publique – Hôpitaux de Paris, Université Paris-Est Créteil, Hôpital Intercommunal de Créteil

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Variable analysis

independent variables

Caspase3 antibody
Caspase9 antibody
Actin antibody
PhosphoAKT antibody

dependent variables

Expression levels of caspase3, caspase9, actin, and phosphoAKT

control variables

Western blot protocol as previously described [7]
Rabbit monoclonal antibodies (1/1000 dilution)
Secondary polyclonal mouse anti-rabbit antibody HRP conjugated (1/2000 dilution)
HRP and chemiluminescent peroxidase substrate (#CPS1120, Sigma) for blot development
Data collection using Geliance CCD camera (Perkin Elmer)
Data analysis using ImageJ software (NIH)

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