PCR primers were designed to amplify, a 579 bases 5’ segment of GLI1AS, a 407 bases 3’ segment of GLI1AS and a full-length GLI1AS from cDNA (Supplementary figure 1A ), and the products were cloned into the KpnI and BamHI sites of the pCMV5 vector. The GLI1 expression construct (Flag-tagged) has been described previously (Shimokawa et al., 2008 (link)). The constructs were used to transfect Rh36 cells as described before (Tostar et al., 2012 (link)). Western analysis was performed using a GLI1 rabbit polyclonal antibody (Cell Signaling) and a Vinculin mouse monoclonal antibody (Sigma-Aldrich) as previously (Tostar et al., 2012 (link)).
Vinculin mouse monoclonal antibody
Manufactured by Merck Group
Vinculin mouse monoclonal antibody is a laboratory reagent used to detect the presence and distribution of the vinculin protein in biological samples. Vinculin is a cytoskeletal protein involved in cell-cell and cell-matrix adhesion. This antibody can be used in various immunodetection techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to study the localization and expression of vinculin in different cell types and tissues.
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2 protocols using vinculin mouse monoclonal antibody
1
Cloning and Transfection of GLI1AS Constructs
2
Cloning and Transfection of GLI1AS
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PCR primers were designed to amplify, a 579 bases 5′ segment of GLI1AS, a 407 bases 3′ segment of GLI1AS and a full‐length GLI1AS from cDNA (Supplementary Figure 1A ), and the products were cloned into the KpnI and BamHI sites of the pCMV5 vector. The GLI1 expression construct (Flag‐tagged) has been described previously (Shimokawa et al., 2008 ). The constructs were used to transfect Rh36 cells as described before (Tostar et al., 2012 ). Western analysis was performed using a GLI1 rabbit polyclonal antibody (Cell Signaling) and a Vinculin mouse monoclonal antibody (Sigma–Aldrich) as previously (Tostar et al., 2012 ).
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