EpiTriton Histone Peptide Array Version 1.0

Covering 296 single and combinatorial modifications on histone H3, H4, H2A, or H2B

User Manual

Product Description

  • Examine the selectivity and specificity of histone modification antibodies
  • Analyze the specificity of histone binding proteins
  • Identify substrates for histone-modifying enzymes

EpiCypher's EpiTriton™ Histone Peptide Array platform is designed for rapid and high-throughput screening of effector protein, antibody and enzyme interactions with a comprehensive library of combinatorially-modified and biotinylated histone peptides immobilized on a streptavidin-coated glass slide. The peptides encompass over 296 unique modifications on the four core histones (H2A, H2B, H3 and H4) and several histone variants. Every EpiTriton™ Histone Peptide Array contains more than 292 histone peptides spotted 12 times each for high quality detection and analysis of antibody or protein binding or enzyme activity. Extensive validation and purification of peptides is performed prior to histone peptide array printing to ensure the highest quality product for your studies.

  • Unmatched quality and diversity
  • Industry leading reproducibility (each batch controlled by a fluorescent tracer co-spotted with each peptide and performance validated using a histone binding effector protein with a known binding pattern)
  • Highly pure modified histone peptides (Quality assured by HPLC and Mass Spec)
  • Positive controls for epitope tags and primary antibodies included
  • Three subarrays per slide, multichamber gasket for parallel experiments included
  • Extensive PTM coverage incl. methylation, acetylation, phosphorylation as well as acyl family modifications (e.g. crotonyl, butyryl etc.)
  • Biotinylated peptides also available as individual products

EpiTriton™ Histone Peptide Array Design

Epitriton Microarray Layout 700

Fig.1. Each EpiCypher EpiTriton™ Histone Peptide Array contains 296 biotinylated histone peptides (20 amino acids in length or more) immobilized on a streptavidin-coated glass slide. Peptides are spotted as three identical sub-arrays, labeled A, B and C. Within each sub-array, each peptide is spotted twice in groups of 3 (red dots), for a total of 18 spots per peptide on each slide.

EpiTriton Histone Peptide Array typical workflow

Epitriton Workflow 750 2

Fig.2. For detection of the interaction of an effector protein with peptides on the array as shown above, you need a primary antibody to the protein (or to an affinity tag) and a fluorescently-labeled (or HRP-conjugated for ECL detection) secondary antibody to the primary. This is much like the detection procedure employed in immunofluorescence miscroscopy. For analysis of histone antibody specificity, you need a primary antibody to a histone modification and a labeled (fluorescent or HRP-conjugated) secondary antibody recognizing the primary antibody.

EpiTriton™ Histone Peptide Array Data: Histone Methyltransferase Enzyme Assay

Kmt Array Data 700

Fig.3. A recombinant histone lysine 9 methyltransferase was used to methylate peptides on the EpiTriton™Histone Peptide Array. Left Panel (-KMT): No enzyme control. Right panel (+KMT): Enzyme applied to array.
Methylation was detected using an antibody recognizing H3K9me2. White boxes highlight H3K9 methylated peptides detected by the antibody prior to enzyme addition. New spots detected depict novel sites of H3K9me2 on the array. These results highlight the use of the EpiTriton™ Histone Peptide Array for use in detecting the substrate specificity of histone modifying enzymes.

Supplemental Files

EpiTriton™ Annotated Peptide List v5 

EpiTriton™ Annotated Peptide Sequences

Related Products

Individual Histone Modification Peptides


Rothbart SB et al (2015). An Interactive Database for the Assessment of Histone Antibody Specificity. Mol Cell 59: 502-511. LINK

Ali M et al (2015). Molecular insight into inhibition of the methylated histone-plant homeodomain complexes by calixarenes. J Biol ChemLINK

Fuchs SM, Krajewski K, Baker RW, Miller VL, Strahl BD (2011). Influence of combinatorial histone modifications on antibody and effector protein recognition. Curr Biol 21: 53-58. Link

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Dr. Sieke Schaepe

Tel. +49 (0) 6221 71415 16 info@biocat.com