Understanding epigenetic reader proteins and their binding preference for post-translational modifications (PTMs) is key to unveiling how these proteins regulate genome processes and how their dysregulation may be contributing to disease. The Captify™ Nucleosome Panel enables access to epigenetic diversity in a physiologically relevant context, the nucleosome, where chromatin reader activity can be accurately interrogated. The 96-well plate contains recombinant mononucleosomes as well as appropriate controls. Human histones expressed in E. coli bearing single and combinatorial histone post-translational modifications (PTMs) are wrapped by 147 or 199 base pair DNA with a 5’ biotin-TEG group and a central 601-positioning sequence, identified by Lowary and Widom
All nucleosomes in the panel are subjected to EpiCypher’s rigorous quality control metrics, including: ESI-TOF mass spectrometry analysis of the modified histones, SDS-PAGE to confirm octamer composition and purity, native PAGE to confirm nucleosome assembly, and western blot analysis of the PTM, histone mutation, or histone variant (if applicable).
Specifications:
Storage: Stable for six months at -80°C from date of receipt. Avoid freeze/thaws.
Formulation: The material included in this 96-well plate is provided at 1.5 µM in 5 µL of nucleosome storage buffer (10 mM Tris-HCl pH 7.5, 25 mM NaCl, 1 mM EDTA, 2 mM DTT*, & 20% glycerol). *dNucs containing crotonyl-lysine are stored in nucleosome storage buffer without DTT. MW = ~200,000 Da
Gene & Protein Information: Uniprot ID:
H2A – P04908 (alt. names: H2A type 1-B/E, H2A.2, H2A/a, H2A/m)
H2B – O60814 (alt. names: H2B K, HIRA-interacting protein 1)
H3.1 – P68431 (alt. names: H3, H3/a, H3/b, H3/c, H3/d)
H3.2 – Q71DI3
H3.3 – P84243
H4 – P62805
Application Notes: Access to epigenetic diversity in the context of a physiological nucleosome enables broad end-user applications, including nucleosome binding studies (e.g. chromatin reader binding preferences [2-4] see Figure 2), enzyme screening assays (e.g. identification of preferred substrates), and antibody specificity testing (e.g. for applications where histone peptide specificity is an insufficient surrogate). The biotin group on the DNA facilitates a wide variety of applications involving streptavidin capture.
