Immunofluorescence Analysis Protocol

Immunofluorescence analysis was carried out as previously described (Sabbattini et al., 2007 (link)). Secondary antibodies used for fluorescence detection were Alexa 488–anti-rabbit and Alexa 488–anti-rat (both from Molecular Probes, Eugene, OR). Images were collected by confocal microscopy using a SP5 microscope (Leica Microsystems, Wetzlar, Germany) and LAS-AF software.

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Sabbattini P., Sjoberg M., Nikic S., Frangini A., Holmqvist P.H., Kunowska N., Carroll T., Brookes E., Arthur S.J., Pombo A, & Dillon N. (2014). An H3K9/S10 methyl-phospho switch modulates Polycomb and Pol II binding at repressed genes during differentiation. Molecular Biology of the Cell, 25(6), 904-915.

Publication 2014

Alexa 488–anti-rat SP5 microscope

Antibodies Confocal microscopy Fluorescence Immunofluorescence Microscope Molecular probes Rabbit

Corresponding Organization :

Other organizations : Hammersmith Hospital, MRC Protein Phosphorylation and Ubiquitylation Unit, University of Dundee

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Fluorescence detection

control variables

Not explicitly mentioned

controls

Positive control: Not specified
Negative control: Not specified

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