Selected article for: "ice cold and room temperature"
Author: Kang, Na-Jin; Koo, Dong-Hwan; Kang, Gyeoung-Jin; Han, Sang-Chul; Lee, Bang-Won; Koh, Young-Sang; Hyun, Jin-Won; Lee, Nam-Ho; Ko, Mi-Hee; Kang, Hee-Kyoung; Yoo, Eun-Sook
Title: Dieckol, a Component of Ecklonia cava, Suppresses the Production of MDC/CCL22 via Down-Regulating STAT1 Pathway in Interferon-? Stimulated HaCaT Human Keratinocytes
  • Document date: 2015_5_1
    • ID: k6f3luil_16
    Snippet: HaCaT cells were seeded onto round coverslips in a 6-well plate and incubated were stimulated with IFN-γ in the presence of dieckol for 24 h. Fixed with freshly prepared 3.7% paraformaldehyde for 30 min, and permeabilized with ice-cold 100% MeOH for 10 min at -20°C. After a 1 h incubation with 3% BSA/0.1% Triton X-100/PBS, the cells were incubated with primary anti-STAT1 antibodies overnight at 4 o C. The cells were washed and then incubated wi.....
    Document: HaCaT cells were seeded onto round coverslips in a 6-well plate and incubated were stimulated with IFN-γ in the presence of dieckol for 24 h. Fixed with freshly prepared 3.7% paraformaldehyde for 30 min, and permeabilized with ice-cold 100% MeOH for 10 min at -20°C. After a 1 h incubation with 3% BSA/0.1% Triton X-100/PBS, the cells were incubated with primary anti-STAT1 antibodies overnight at 4 o C. The cells were washed and then incubated with DyLight488-conjugated donkey anti-rabbit (BioLegend, San Diego, CA, USA) secondary antibody for 30 min at room temperature. After several additional washing steps, the coverslips were mounted in VECTASHIELD mounting media with DAPI (Vector Labs, Burlingame, CA, USA). Fluorescently labeled STAT1 was visualized by using a FV500 confocal microscope (Olympus Corp., Tokyo, Japan).

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