Immunofluorescence Staining of β-Catenin

Immunofluorescence staining was performed as described [30] (link), [40] (link), [43] (link), [50] , [53] (link), [54] (link). Briefly, cells were infected with AdWnt3A or AdGFP for 48 h, fixed with methanol, permeabilized with 1% NP-40, and blocked with 10% BSA, followed by incubating with β-catenin antibody (Santa Cruz Biotechnology). After being washed, cells were incubated with Texas Red-labeled secondary antibody (Santa Cruz Biotechnology). Stains were examined under a fluorescence microscope. Stains without primary antibodies, or with control IgG, were used as negative controls.

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Deng F., Chen X., Liao Z., Yan Z., Wang Z., Deng Y., Zhang Q., Zhang Z., Ye J., Qiao M., Li R., Denduluri S., Wang J., Wei Q., Li M., Geng N., Zhao L., Zhou G., Zhang P., Luu H.H., Haydon R.C., Reid R.R., Yang T, & He T.C. (2014). A Simplified and Versatile System for the Simultaneous Expression of Multiple siRNAs in Mammalian Cells Using Gibson DNA Assembly. PLoS ONE, 9(11), e113064.

Publication 2014

β-catenin antibody Texas Red-labeled secondary antibody

Antibodies Antibody Cells Fluorescence microscope Immunofluorescence Methanol Np 40 Stains β catenin

Corresponding Organization :

Other organizations : University of Chicago, Army Medical University

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Variable analysis

independent variables

AdWnt3A infection
AdGFP infection

dependent variables

β-catenin localization/expression

control variables

Cell type
Methanol fixation
NP-40 permeabilization
BSA blocking
Primary antibody (β-catenin)
Secondary antibody (Texas Red-labeled)

controls

Negative controls: Stains without primary antibodies, or with control IgG

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