Immunocytochemistry

SK Seongjae Kim
JP Jung Min Park
SP Soeun Park
EJ Eunsun Jung
DK Dongmi Ko
MP Minsu Park
JS Juyeon Seo
KN Kee Dal Nam
YK Yong Koo Kang
KL Kyoungmin Lee
LF Lee Farrand
YK Yoon-Jae Kim
JK Ji Young Kim
JS Jae Hong Seo
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Immunocytochemistry was conducted to assess the expression and co-localization of EGFR and CD44 in MDA-MB-231 cells. Cells in Falcon® chambered cell culture slides (BD Biosciences) were fixed with 4% paraformaldehyde, washed with PBS, and incubated with 0.2% Triton X-100 for 13 min. Primary antibodies were applied to the cells in antibody-diluent (Dako, Denmark) were incubated overnight at 4°C. For secondary antibody reactions, Alexa Fluor®-488 or -594 conjugated secondary antibodies (Thermo Fisher Scientific) were used for staining and then mounted with ProLong® Gold Antifade Reagent with DAPI (Thermo Fisher Scientific). Imaging of the cells was performed using a Carl Zeiss confocal microscope (Weimar, Germany), and the intensity of the EGFR and CD44 signal was analyzed using the intensity profile tool.

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