Immunohistochemical Visualization of Microtubules and Actin

Immunohistochemistry was performed as described previously [5 (link)]. For this study, we used combined stainings of anti-α-tubulin or anti-serotonin with phalloidin-rhodamine. For anti-α-tubulin staining, a mixture of anti-acetylated α-tubulin (monoclonal anti-tubulin, acetylated antibody, produced in mouse, ascites fluid, Sigma-Aldrich, St. Louis, MO, USA; dilution 1:500 in PBST-NGS) and anti-tyrosinated α-tubulin (monoclonal anti-tubulin, tyrosine antibody, produced in mouse, ascites fluid, Sigma-Aldrich, St. Louis, MO, USA; dilution 1:250 in PBST-NGS) was used. The labeled α-tubulin is a structural component of microtubules, which are amongst others present in axons, while serotonin (=5-HT) is a neurotransmitter. Phalloidin-rhodamine labels filamentous muscular actin (f-actin) [5 (link)].

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Weidhase M., Beckers P., Bleidorn C, & Aguado M.T. (2016). On the role of the proventricle region in reproduction and regeneration in Typosyllis antoni (Annelida: Syllidae). BMC Evolutionary Biology, 16, 196.

Publication 2016

monoclonal anti-tubulin, acetylated antibody anti-tyrosinated α-tubulin

Antibody Ascites fluid Axons Dilution F actin Immunohistochemistry Microtubules Mouse Muscular Neurotransmitter Phalloidin Rhodamine Rhodamine phalloidin Tubulin Tyrosine α tubulin

Corresponding Organization :

Other organizations : Leipzig University, University of Bonn, Autonomous University of Madrid

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Variable analysis

independent variables

Staining type (anti-α-tubulin, anti-serotonin, phalloidin-rhodamine)

dependent variables

Localization/distribution of microtubules, serotonin, and filamentous actin

control variables

Experimental procedure (as described previously in reference 5)
Antibody dilution (1:500 for anti-acetylated α-tubulin, 1:250 for anti-tyrosinated α-tubulin, in PBST-NGS)

controls

Positive control: Not specified
Negative control: Not specified

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