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The AlphaLISA technology allows performing no-wash homogeneous, proximity immunoassays using Alpha Donor and AlphaLISA, Acceptor beads. In this technical note, we present an optimized, assay for measuring changes in the levels of H3K27ac after treatment, of cells with sodium butyrate and Trichostatin A (TSA), two nonselective, histone deacetylase (HDAC) inhibitors. Following a, homogeneous histone extraction protocol, the mark of interest, is detected by the addition of a biotinylated anti-Histone H3, (C-terminus) antibody and AlphaLISA Acceptor beads conjugated, to an antibody (Ab) specific to the mark. The biotinylated antibody is, then captured by Streptavidin (SA) Donor beads, bringing the two, beads into proximity. Upon laser irradiation of the Donor beads, at 680 nm, short-lived singlet oxygen molecules produced by the, Donor beads can reach the Acceptor beads in proximity to generate, an amplified chemiluminescent signal at 615 nm.
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