Mouse IgG negative control for ChIP
AS21 4695 | Clonality: Monoclonal | Host: Mouse | Reactivity: Negative control IgG for chromatin immunoprecipiation assays (ChIP)

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Reactivity
Application examples

ChIP assay was done with rabbit polyclonal antibody against H3K27ac using chromatin from sheared 1 million HeLa cells. Mouse IgG (AS21 4695) served as a negative IP control. Amount of antibody was 1 μg/ChIP experiment. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. The graph shows recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Detection of H3K27ac on HeLa cells were stained with the primary antibody agains H3K27ac (top) and with DAPI. Mouse control IgG (AS21 4695) was used as a negative control (bottom row).
Fixation: 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA.
Primary antibody: anti-H3K27ac or mouse IgG negative control antibody (left) diluted 1:500 in a blocking solution
Secondary antibody: anti-rabbit antibody conjugated to Alexa594.
Middle panel: staining of the nuclei with DAPI.
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