ChIP was performed as described in Fernius et al. (2013) (link) using mouse anti-HA (12CA5, Roche Diagnostics), mouse anti-PK(V5) (AbD Serotec), or rabbit anti-GFP (a kind gift of Dr. Eric Schirmer, University of Edinburgh) antibodies. For experiments in Supplemental Figure S2, A–C, qPCR was performed using a Bio-Rad iCycler machine and the protocol described in Fernius and Marston (2009) (link). For all other experiments shown, qPCR was performed on a Roche LightCycler.
Mouse anti pk v5
Manufactured by Bio-Rad
Mouse anti-PK(V5) is a monoclonal antibody that specifically recognizes the PK(V5) epitope tag. The antibody is designed for the detection and purification of recombinant proteins fused with the PK(V5) tag.
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2 protocols using mouse anti pk v5
1
ChIP Protocol using Antibodies
2
Western Blotting with Fluorescence Detection
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Samples were prepared for Western blotting as described in Clift et al. (2009) (link) except that some antibodies were detected using the fluorescence-based Li-Cor Odyssey system. Antibodies used were mouse anti-HA 12CA5 (Roche), mouse anti-PK(V5) (AbD Serotec), mouse anti-aid (Cosmo Bio Co.), and rabbit or mouse anti-Pgk1 (laboratory stock and Life Technologies, respectively).
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