Antibody-Based Immunofluorescence and Immunoblot Analysis

Monoclonal antibodies GFP 3E1 (Cancer Research UK), Actin AC-74 (Sigma), and zyxin 164D4 (SySy) as well as rabbit antibodies against Abi1 (Sigma A5106) and VASP (Cell Signaling 3132S) were used. Antibodies against human Abi1, Nap1, and PIR121 (Steffen et al., 2004 (link)) were kindly provided by Dr Theresia Stradal (University of Muenster, Germany). Antibodies against Drosophila Abi and WAVE have been described (Bogdan et al., 2005 (link), Stephan et al., 2011 (link)). Immunofluorescence and immunoblot analyses were performed as described (Boëda et al., 2007 (link)). Projection patterns of all photoreceptor axons of the indicated genotypes were depicted using mouse anti-24B10 (α-Chaoptin, DSHB; Stephan et al., 2011 (link)).

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Chen X.J., Squarr A.J., Stephan R., Chen B., Higgins T.E., Barry D.J., Martin M.C., Rosen M.K., Bogdan S, & Way M. (2014). Ena/VASP Proteins Cooperate with the WAVE Complex to Regulate the Actin Cytoskeleton. Developmental Cell, 30(5), 569-584.

Publication 2014

Actin AC-74

Abi1 Actin Antibodies Axons Cancer Drosophila Genotypes Human Immunoblot analyses Immunofluorescence Monoclonal antibodies Mouse Nap1 Photoreceptor Rabbit Vasp Zyxin

Corresponding Organization :

Other organizations : The Honourable Society of Lincoln's Inn, Cancer Research UK, University of Münster, The University of Texas Southwestern Medical Center

Top 5 similar protocols

Variable analysis

independent variables

Abi1 antibodies
Nap1 antibodies
PIR121 antibodies
Drosophila Abi antibodies
Drosophila WAVE antibodies

dependent variables

Immunofluorescence analysis
Immunoblot analysis
Photoreceptor axon projection patterns

control variables

Monoclonal antibodies: GFP 3E1, Actin AC-74, zyxin 164D4
Rabbit antibodies: Abi1, VASP

controls

Positive controls: Not explicitly mentioned.
Negative controls: Not explicitly mentioned.

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