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AlphaLISA Acetylated Histone H3 Lysine 9 (H3K9ac) Cellular Detection Kit, 500 Assay Points

This kit is designed for the detection of acetylated Histone H3 Lysine 9 (H3K9ac) in cell lysates using a homogeneous AlphaLISA assay (no wash steps).

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For research use only. Not for use in diagnostic procedures.

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AL714HV
100 assay points
695.00 USD
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AL714C
500 assay points
2159.00 USD
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AL714F
5,000 assay points
12400.00 USD
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Detail Information

The AlphaLISA® Epigenetic Cellular Detection Kits enable rapid and direct detection of endogenous modification of epigenetic marks on histone H3. These no wash, all-in-one-well assays are optimized for simplicity and throughput.

  • Simple assay protocol amenable to automation, eliminating tedious ELISAs and Westerns
  • Suitable for endogenous and recombinant cell lines, providing the flexibility to work with relevant cell models
  • Ideal for HTS protocols with demonstrated Z’ factors greater than 0.6

The AlphaLISA detection of epigenetic marks in cellular extracts is performed as follows: cell cultured in the presence of compounds are lysed with the Cell-Histone Lysis buffer. Histones are then extracted from the nucleosomes by the addition of the Cell-Histone Extraction buffer. AlphaLISA anti-mark Acceptor beads and Biotinylated anti-Histone H3 (C-terminus) antibodies are then added for the capture of histone proteins carrying the mark of interest. After incubation, Streptavidin Donor beads are added for the capture of the biotinylated antibody. In the presence of histone proteins bearing the mark of interest, the beads come into proximity. Excitation of the Donor beads provokes the release of singlet oxygen molecules that trigger a cascade of energy transfer reactions in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.

Specifications

Assay Target Histone H3
Assay Target Class Histone
Automation Compatible Yes
Detection Method Alpha
Experimental Type In vitro
Molecular Modification Acetylation
Product Brand Name AlphaLISA
Shipping Condition Blue Ice
Therapeutic Area Epigenetics
Unit Size 500 assay points
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Brochure

Technical Note

Technical note: AlphaLISA (H3K9ac) Cell Based Kit

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 H3K9ac after treatment of cells, with sodium butyrate and Trichostatin A (TSA), two non-selective, 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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