Comprehensive Protein Expression Analysis

Western blot were carried out as previously described [38 (link)]. Immunostaining was carried out using a goat monoclonal antibodies against RhoA (2117), Rock2 (9029), Rac1 (2465), Cdc42 (2466), SDF-1α (3740), integrin (α4-4600; α5-4705; αV-4711; β3-4702; β4-4707; β5-4708), actin (3200) (1/1000, Cell signaling) and a secondary polyclonal mouse anti-goat antibody HRP conjugated (1/2000, cell signaling). 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., Abu-Kaoud N., Abdesselem H., Madani A., Hoarau-Véchot J., Thawadi H.A., Vidal F., Couderc B., Favre G, & Rafii A. (2015). SDF-1alpha concentration dependent modulation of RhoA and Rac1 modifies breast cancer and stromal cells interaction. BMC Cancer, 15, 569.

Publication 2015

CPS1120 Geliance CCD camera

Actin Anti antibody Cdc42 Goat Integrin Monoclonal antibodies Mouse Peroxidase Rhoa Rock2 Western blot

Corresponding Organization :

Other organizations : Weill Cornell Medical College in Qatar, Cornell University, Institut Claudius Regaud, Centre de Recherche en Cancérologie de Toulouse

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Variable analysis

independent variables

Immunostaining using goat monoclonal antibodies against RhoA, Rock2, Rac1, Cdc42, SDF-1α, integrin (α4, α5, αV, β3, β4, β5), and actin

dependent variables

Protein expression levels of RhoA, Rock2, Rac1, Cdc42, SDF-1α, integrins, and actin

control variables

Western blot protocol (previously described in reference 38)
Concentration of primary antibodies (1/1000)
Concentration of secondary antibody (1/2000)
Chemiluminescent peroxidase substrate used for blot development (#CPS1120, Sigma)
Data collection using Geliance CCD camera (Perkin Elmer)
Data analysis using ImageJ software (NIH)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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