Immunofluorescence Cell Staining Protocol

Immunofluorescence cell staining was performed as previously described (Izumi et al. 2014 (link)). Briefly, cells were fixed with 4% formaldehyde in PBS, permeabilized with 0.2% TritonX-100 in PBS, and blocked with 2% bovine serum albumin in PBS. The cells were incubated with the primary antibodies, followed by incubation in the presence of Alexa488-, Alexa568-, or Cy5-labeled secondary antibody (Thermo Fisher Scientific). The antibodies used in this study are described in Supplemental Table S1.

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Machitani M., Taniguchi I., McCloskey A., Suzuki T, & Ohno M. (2020). The RNA transport factor PHAX is required for proper histone H2AX expression and DNA damage response. RNA, 26(11), 1716-1725.

Publication 2020

Alexa488 Alexa568 Cy5-labeled secondary antibody

Antibodies Antibody Bovine serum albumin Cells Formaldehyde Immunofluorescence

Corresponding Organization :

Other organizations : Kyoto University

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Variable analysis

independent variables

Fixation (4% formaldehyde in PBS)
Permeabilization (0.2% TritonX-100 in PBS)
Blocking (2% bovine serum albumin in PBS)
Primary antibodies
Secondary antibodies (Alexa488-, Alexa568-, or Cy5-labeled)

dependent variables

Antibody localization/binding (measured by immunofluorescence)

control variables

PBS (for fixation, permeabilization, and blocking)
Cell type (not explicitly mentioned)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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