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Histone PTM Multiplex Service

profile histone modifications in multiplex

Active Motif's Histone PTM Quantitation Service gives you access to ground-breaking technology that enables high-throughput multiplexed screening of histone modifications. The service uses Luminex® xMAP® magnetic bead-based multiplexing technology to simultaneously measure multiple histone modification targets in a single reaction. This first-of-its-kind multiplex epigenetic assay generates more data with less sample than traditional western blot or ELISA methods.

 

Technology covered under U.S. Patent No. 8367334 b2.

Available modifications for multiplex analysis.
Available modifications for multiplex analysis

(Click image to enlarge)

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Assay Features

  • Minimal cell requirement of 250,000 cells is sufficient to quantify 13 histone modifications.
  • Detects ≥ 10% difference between samples.
  • Controls include total H3 and samples containing known PTM changes.

Histone PTM Quantitation Service includes:

  • Assay design.
  • Sample preparation.
  • Completion of the Luminex assay.
  • Data analysis.
Histone Services Protocol
Luminex Cell Prep Protocol form

Histone PTM
Cell Prep Protocol
form

Custom Services Brochure

Custom Services
 brochure

 
Name Cat No. Price  
Histone PTM Quantitation, 8 Sample Service 25062 Get Quote
Luminex Histone PTM Bead Set 25063 Get Quote
Active Motif Epigenetic Services Brochure
Newsletter - October 2015
The Histone H3 PTM Multiplex Assay shows increased histone acetylation in response to SAHA-mediated HDAC inhibition
Figure 1: Data from our Histone PTM Quantitation Service shows increased histone acetylation in response to SAHA-mediated HDAC inhibition (Click image to enlarge)

HeLa cells treated with increasing concentrations of the HDAC inhibitor SAHA were evaluated in a multiplex of H3 pan-acetyl, H3S10ph, H3K9ac, H3K4me3, H3K27me2 & H3 Total Ab-conjugated beads using the Histone H3 PTM Multiplex Assay. Total H3 beads were used for normalization. The results demonstrate the ability of the assay to simultaneously assess specific and off-target effects of the treatment on histone modification levels. The dashed lines represent IC50 values, 4.0 μM and 4.6 μM, determined for pan-acetyl and H3K9ac, respectively.

 

EZH2 inhibition results in decreased H2K27me3 levels
Figure 2: HDAC inhibitor (VPA) treatment increases histone acetylation with concomitant reductions in H3K27me2

HeLa cells treated with increasing concentrations of the HDAC inhibitor VPA were evaluated by Luminex in a multiplex of H3 pan-acetyl, H3S10ph, H3K9ac, H3K27me2, and H3 Total Ab-conjugated beads. Total H3 beads were used for normalization. The results demonstrate the ability of the Histone PTM Quantitation Service to detect a dose dependent increase in total and H3K9 specific acetylation. A decrease in H3K27me2 is also observed.

 

HDAC inhibitor (VPA) treatment increases histone acetylation with concomitant reductions in H3K27me2
Figure 3: EZH2 inhibition results in decreased H2K27me3 levels

HeLa cells treated with increasing concentrations of the EZH2 inhibitor GSK126 for 8 days were evaluated by Luminex in a multiplex of H3K9me3, H3K27me3, and H3 Total Ab-conjugated beads. Total H3 beads were used for normalization. EZH2 is a histone H3 lysine 27 specific methyltransferase and inhibition of this enzyme is expected to result in reduced H3K27me3 levels. The results show reduced H3K27me3 at all drug concentrations with no affect on H3K9me3.