ELISA (recommended work dilution= 1:5000-8000 (Only recognized asymmetric form)), Western Blotting (recommended work dilution= 1-2 µg/ml)
Designed and synthetic peptide containing H3R2a was used as immunogen. This antibody was obtained from a mice immunized with the immunogen, then fusion, selection and identification. This antibody was produced from a hybridoma resulting from the fusion of a myeloma with B cells deriveded from the immunized mouse. The IgG fraction of cell culture supernatant was purified by Protein G affinity chromatography.
A separate vial of dilution buffer is provided for reconstitution. The antibody is supplied lyophilized, originally containing PBS, without preservative stabilizers (e.g. sodium azide). The final amount is indicated on the shipping vial.
Histones are highly conserved and can be grouped into five major classes: H1/H5, H2A, H2B, H3, and H4. The tight packing of DNA into chromatin creates a need for mechanisms to relax chromatin and expose DNA for transcription, replication, and DNA repair. One of the mechanisms used by the cell to access DNA is the post-translational modification of Histone tails. Specifically, methylation of Histone tails generates a docking site for effector proteins, which aid in the recruitment of other enzymes necessary for the function at hand. Methylation at lysine and arginine residues within Histones has been linked to gene expression. In general, methylation of Histone residues lysine 4 and 36 on H3 are correlated with active gene regions, while methylation of lysine 9 and 27 on H3 is correlated with repressed gene regions. Very importantly, methylation of Histone residues arginie 2 on H3 (termed as H3R2) involved in gene expression.
The antibody is stable for at least 1 year from the date of receipt when stored at -20°C to -70°C. Reconstituted antibody can also be aliquotted and stored at 4°C for 1 month or at -20°C to -70°C in a manual defrost freezer for many months without detectable loss activity. Please avoid freeze-thaw cycles.
The antibody is not only able to bind to H3R2a, but also distinguish two forms (Asymmetric and Symmetric) of H3R2. It specifically detected protein derived from cell lysates in both ELISA and WB.
- Guccione E, Bassi C, Casadio F, et al. Methylation of histone H3R2 by PRMT6 and H3K4 by an MLL complex are mutually exclusive. Nature 2007;449:933937.
- Barski A, Cuddapah S, Cui K, et al. High-resolution profiling of histone methylations in the human genome. Cell. 2007;129:823837.