Selected article for: "primary antibody and room temperature"
Author: Zhou, Jie; Li, Cun; Sachs, Norman; Chiu, Man Chun; Wong, Bosco Ho-Yin; Chu, Hin; Poon, Vincent Kwok-Man; Wang, Dong; Zhao, Xiaoyu; Wen, Lei; Song, Wenjun; Yuan, Shuofeng; Wong, Kenneth Kak-Yuen; Chan, Jasper Fuk-Woo; To, Kelvin Kai-Wang; Chen, Honglin; Clevers, Hans; Yuen, Kwok-Yung
Title: Differentiated human airway organoids to assess infectivity of emerging influenza virus
  • Document date: 2018_6_26
    • ID: z637eh2z_25
    Snippet: Immunofluorescence Staining. To identify the indicated cell types and the virusinfected cells, immunofluorescence staining was applied to the 3D and 2D AOs. Briefly, the organoids were fixed with 4% PFA, permeabilized with 0.1-5% Triton X-100, and blocked with Protein Block (Dako). Then the organoids were incubated with primary antibodies (SI Appendix, Table S3 ) diluted in Antibody Diluent buffer (Dako) overnight at 4°C, followed by incubation .....
    Document: Immunofluorescence Staining. To identify the indicated cell types and the virusinfected cells, immunofluorescence staining was applied to the 3D and 2D AOs. Briefly, the organoids were fixed with 4% PFA, permeabilized with 0.1-5% Triton X-100, and blocked with Protein Block (Dako). Then the organoids were incubated with primary antibodies (SI Appendix, Table S3 ) diluted in Antibody Diluent buffer (Dako) overnight at 4°C, followed by incubation with secondary antibodies (SI Appendix, Table S3 ) for 1-2 h at room temperature. Nuclei and actin filaments were counterstained with DAPI (Invitrogen) and Phalloidin-647 (Sigma Aldrich), respectively. The confocal images were acquired using a Carl Zeiss LSM 780 or 800 confocal microscope.

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