Following the addition of vehicle control or COMET probes (BG14, BG47, BG48) at a final concentration of 25 μM, MCF-7 cells were exposed to varying amounts of 470 nm light (8.5 mW/cm2) for 16 h in a tissue culture incubator (37 °C, 5% CO2). The light exposure was modulated at 1 Hz (no light, 250 ms on/750 ms off, 750 ms on/250 ms off). Subsequently, cells were lysed and the relative amount of acetylated H3K9 was analyzed by standard western blot analysis (rabbit anti-H3K9ac antibody, Millipore #07-352 1:10,000, anti-rabbit-horseradish peroxidase (HRP) 1:2,000, Cell Signaling #7074) using anti β-Actin as loading control (Sigma A1978, anti-mouse-HRP Cell Signaling #7076 1:10,000). Fold increase over DMSO treated samples were quantified with ImageJ software (http://imagej.nih.gov/ij ). Western blot assays were performed in triplicate with each treatment done in duplicate. Differences in mean acetylation levels comparing presence and absence of light were assessed using a two-sided t-test with p<0.05 considered significant.
Rabbit anti h3k9ac antibody
Manufactured by Merck Group
The Rabbit anti-H3K9ac antibody is a laboratory reagent used to detect and analyze the acetylation of lysine 9 on histone H3 proteins. It is a specific and sensitive tool for researchers studying epigenetic mechanisms and chromatin modification in various biological systems.
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Lab products found in correlation
2 protocols using rabbit anti h3k9ac antibody
1
Light-Induced Histone Acetylation Assay
2
Light-Induced Histone Acetylation Assay
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Following the addition of vehicle control or COMET probes (BG14, BG47, BG48) at a final concentration of 25 μM, MCF-7 cells were exposed to varying amounts of 470 nm light (8.5 mW/cm2) for 16 h in a tissue culture incubator (37 °C, 5% CO2). The light exposure was modulated at 1 Hz (no light, 250 ms on/750 ms off, 750 ms on/250 ms off). Subsequently, cells were lysed and the relative amount of acetylated H3K9 was analyzed by standard western blot analysis (rabbit anti-H3K9ac antibody, Millipore #07-352 1:10,000, anti-rabbit-horseradish peroxidase (HRP) 1:2,000, Cell Signaling #7074) using anti β-Actin as loading control (Sigma A1978, anti-mouse-HRP Cell Signaling #7076 1:10,000). Fold increase over DMSO treated samples were quantified with ImageJ software (http://imagej.nih.gov/ij ). Western blot assays were performed in triplicate with each treatment done in duplicate. Differences in mean acetylation levels comparing presence and absence of light were assessed using a two-sided t-test with p<0.05 considered significant.
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