Total Histone H3 ELISA

Total Histone H3 ELISA

Histone modifications such as phosphorylation, acetylation and methylation at specific amino acid residues on the histone tails that extend beyond the core nucleosome have been found to influence and regulate transcription, chromosome packaging and DNA damage repair. The Total Histone H3 ELISA makes it easy to screen for changes in histone H3 levels, regardless of the modification state. The assay works with core histone preparations or histones purified by acid extraction from tissue or cell samples (Figure 1).

In addition to screening total histone H3 levels, the Total Histone H3 ELISA can be used to normalize the methylation level of your samples when run in parallel with Active Motif's Histone H3 methylated Lys27 ELISAs. For normalization, the Total Histone H3 Kit and Histone Modification ELISA should be run in parallel. The methylation specific data is then normalized by dividing it by the total histone H3 values.

Figure 1: Total Histone H3 ELISA.
The Total Histone H3 ELISA was used to assay purified HeLa core histones (62.5 - 125 ng) made using Active Motif's Histone Purification Mini Kit (Catalog No. 40026) and HeLa acid extracts (500 ng - 1 µg). The provided Recombinant Histone H3 protein was assayed from 1.56 - 100 ng/well as a reference standard curve. Data shown are the results from wells assayed in duplicate. These results are provided for demonstration only.

The Histone ELISA Method

The Total Histone H3 ELISA kit is a sandwich ELISA for screening the levels of histone H3 in human, mouse and rat systems. The kits utilize a histone H3 monoclonal antibody to capture histone H3 from purified core histones or histones isolated by acid extraction from tissue or cell samples. A C-terminal Total Histone H3 primary antibody is used for detection, while a secondary antibody conjugated to horseradish peroxidase (HRP) and developing solutions provide a sensitive colorimetric readout that is easily quantified by spectrophotometry. The assay is performed in a convenient 96-stripwell plate, enabling you to simultaneously screen from 1 to 96 samples in a single experiment and save valuable time over immunoblotting methods.

Included in each Histone Modification ELISA is a methylated recombinant histone made using Active Motif's patented protein synthesis technology. Each protein has been specifically methylated at the desired lysine site and is >99% pure. This protein can be used to build a reference standard curve to quantitate the amount of specifically methylated histone H3 in your samples. The Total Histone H3 ELISA includes an unmodified Recombinant Histone H3 protein.

Contents & Storage

Total Histone H3 ELISA Kits include a 96-stripwell Histone H3 Capture plate, Total Histone H3 antibody (C-terminal), HRP-conjugated anti-rabbit IgG secondary antibody, Assay Dilution Buffer, 20X Wash Buffer, Developing Solution, Stop Solution, unmodified Recombinant Histone H3 positive control protein and plate sealer. Storage conditions vary from room temperature to -80°C, see manual for details. All reagents are guaranteed stable for 6 months when stored properly.