Immunocytochemistry of Tricellulin and Occludin

For immunocytochemistry, cells were grown on 35-mm glass-base dishes (Iwaki, Chiba, Japan) coated with rat tail collagen. They were fixed with an ethanol and acetone 1:1 mixture at −20 °C for 10 min. Staining was performed as described previously22 (link) using the primary polyclonal anti-tricellulin antibody and monoclonal anti-occludin antibody. The specimens were examined using an epifluorescence microscope (Olympus, Tokyo, Japan) and a confocal laser scanning microscope (LSM510; Carl Zeiss, Jena, Germany).

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Takasawa A., Murata M., Takasawa K., Ono Y., Osanai M., Tanaka S., Nojima M., Kono T., Hirata K., Kojima T, & Sawada N. (2016). Nuclear localization of tricellulin promotes the oncogenic property of pancreatic cancer. Scientific Reports, 6, 33582.

Publication 2016

35-mm glass-base dishes epifluorescence microscope LSM510

Acetone Anti antibody Cells Collagen Confocal laser scanning microscope Dishes Ethanol Immunocytochemistry Microscope Monoclonal antibody Occludin Tail Tricellulin

Corresponding Organization :

Other organizations : Sapporo Medical University, The University of Tokyo

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Variable analysis

independent variables

Coating of 35-mm glass-base dishes with rat tail collagen

dependent variables

Cellular localization of tricellulin and occludin proteins visualized using immunocytochemistry and fluorescence microscopy

control variables

Cell culture conditions (e.g., cell type, growth medium, temperature, time)
Fixation method (ethanol and acetone mixture at -20°C for 10 min)
Primary antibodies used (polyclonal anti-tricellulin and monoclonal anti-occludin)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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