Western blot analysis was performed essentially as previously described (Vucetic et al., 2008 (link); Zhao et al., 2009 (link)). Primary antibodies used were goat anti-GAPDH (sc-48176, Santa Cruz), mouse anti-V5 (R960-25, Life Technologies), mouse anti-FLAG (F1804, Sigma-Aldrich,), mouse anti-H3 histone (10799, Abcam) and rabbit anti-HP1α (2616, Cell Signaling, Beverly, MA). Secondary antibodies used were donkey anti-mouse IRDye 800CW, donkey anti-rabbit IRDYE 800CW, and donkey anti-goat IRDye 680CW purchased from LI-COR, Lincoln, NE. Images were captured and quantitated using the LI-COR Odyssey instrument and software. GAPDH levels were used as the loading control.
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Western Blotting Protein Detection
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Corresponding Organization : Temple University
Other organizations : Chestnut Hill College
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Variable analysis
independent variables
- None explicitly mentioned
- Protein expression levels of GAPDH, V5, FLAG, H3 histone, and HP1α
- GAPDH levels used as loading control
- Positive controls: None mentioned
- Negative controls: None mentioned
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