Whole Mount Staining of Orb2, Bol, and Other Proteins

Whole mount staining was performed as in [29] (link). Antibodies used were as follows: mouse anti-Orb2 2D11 and 4G8 IgG (undiluted, developed in the lab) [29] (link), rabbit anti-Bol (1∶1000, a gift from Steven Wasserman) [46] (link), monoclonal anti-β-Tubulin E7 1∶50 (Developmental Studies Hybridoma Bank), rabbit anti-aPKCζ (1∶1000, clone c-20, sc-216, Santa Cruz Biotechnology), rabbit anti-GFP (1∶1000, Cristea Lab, Princeton University). Rabbit polyclonal anti-Dlg-PDZ1 (1∶1000) and guinea pig anti-Bazooka (1∶500) were provided by Yu-Chiun Wang [47] (link). Actin was stained with Alexa488-phalloidin (Invitrogen, Carlsbad, CA). DNA was stained with Hoechst (1∶1000). Secondary antibodies used were goat anti-mouse IgG Alexa 488, 546 or 647, goat anti-rabbit Alexa 488, 546 or 647 (Molecular Probes, Inc.). Samples were mounted in Aqua-polymount on slides for an inverted Zeiss LSM510 or Leica SP5 confocal microscope.

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Xu S., Tyagi S, & Schedl P. (2014). Spermatid Cyst Polarization in Drosophila Depends upon apkc and the CPEB Family Translational Regulator orb2. PLoS Genetics, 10(5), e1004380.

Publication 2014

rabbit anti-aPKCζ Alexa488-phalloidin LSM510

Actin Anti igg Antibodies Clone Confocal microscope Goat Guinea pig Hybridoma Molecular probes Mouse Phalloidin Rabbit Tubulin

Corresponding Organization :

Other organizations : Princeton University, International Center for Public Health

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Variable analysis

independent variables

Antibodies used, including mouse anti-Orb2 2D11 and 4G8 IgG, rabbit anti-Bol, monoclonal anti-β-Tubulin E7, rabbit anti-aPKCζ, rabbit anti-GFP, rabbit polyclonal anti-Dlg-PDZ1, and guinea pig anti-Bazooka

dependent variables

Not explicitly mentioned

control variables

Hoechst stain for DNA, Alexa488-phalloidin stain for actin
Secondary antibodies used, including goat anti-mouse IgG Alexa 488, 546 or 647, and goat anti-rabbit Alexa 488, 546 or 647

controls

Positive controls: Not specified
Negative controls: Not specified

Annotations

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