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Histone Modification ELISAs provide a simple, sensitive method for detecting changes in histone methylation levels from purified core histones or histones isolated by acid extraction. For added convenience and a more quantitative interpretation of results, the Histone Modification ELISAs also include a site- and degree-specific methylated recombinant histone protein for use as a reference standard curve. For complete details, click the ELISA Method tab below.
Histone H3 lysine 9 methylation (H3K9) has been correlated with heterochromatin formation and transcriptional repression, making the methylation state of lysine 9 an interesting marker of transcriptional activity. Histone H3 methylated Lys9 ELISAs are sandwich ELISAs for screeing the levels of histone H3 dimethyl or trimethyl Lys9 in your samples. The methylation specific histone H3 antibodies will not cross-react with other site- or degree-specific methylated histones, enabling small changes in methylation levels to be detected. Click on the H3K9 Info tab below for data and more information.
| Name | Format | Cat. No. | Price | |
|---|---|---|---|---|
| Histone H3 dimethyl Lys9 ELISA | 1 x 96 rxns | 53108 | $455 |
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| Histone H3 trimethyl Lys9 ELISA | 1 x 96 rxns | 53109 | $455 |
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Histone H3 methylated Lys9 (H3K9)
The methylation of lysine 9 on histone H3 (H3K9) has been correlated with the regulation of gene transcription, making methylated lysine 9 on histone H3 a significant marker in studying the state of transcription activity. Methylation of histone H3 at lysine 9 is linked to heterochromatin formation and transcriptional repression. Histone methyltransferases G9a, G9a-related protein (GLP) and SUV39H1 are believed to have specificity for methylation of lysine 9 on histone H3, but only SUV39H1 is able to recruit the chromodomain-containing heterochromatin protein 1 (HP1) to chromatin.
Histone H3 dimethyl Lys9 ELISA
The Histone H3 dimethyl Lys9 ELISA is capable of detecting the level of histone H3 dimethylated at lysine 9 from core histone preparations or histones purified by acid extraction from tissue or cell samples (Figure 1). The Histone H3 dimethyl Lys9 sandwich ELISA assay is also tested for cross-reactivity with other degree-specific recombinant proteins as shown in Figure 2.
Figure 1: Histone H3 dimethyl Lys9 ELISA (H3K9).
The Histone H3 dimethyl Lys9 ELISA was used to assay purified HeLa core histones (125 - 250 ng) made using Active Motif's Histone Purification Mini Kit (Catalog No. 40026) and HeLa acid extracts (5 - 10 µg). The provided Recombinant Histone H3 dimethyl Lys9 protein was assayed from 7.8125 - 500 ng/well as a reference standard curve. Data shown are the results from wells assayed in duplicate. These results are provided for demonstration only.
Figure 2: Specificity of the Histone H3 dimethyl Lys9 ELISA (H3K9).
Recombinant Histone H3, mono-, di- and trimethyl Lys9 proteins were assayed from 15 ng - 500 ng per well using the Histone H3 dimethyl Lys9 ELISA. These results indicate the specificity of the assay. There is extremely low background from histone H3 and little cross-reactivity for mono- or trimethyl Lys9. This means that small, specific changes in dimethyl Lys9 levels can easily be detected with this kit.
Histone H3 trimethyl Lys9 ELISA
The Histone H3 trimethyl Lys9 ELISA is capable of detecting the level of histone H3 trimethylated at lysine 9 from core histone preparations or histones purified by acid extraction from tissue or cell samples (Figure 3). The Histone H3 trimethyl Lys9 sandwich ELISA assay is also tested for cross-reactivity with other degree-specific recombinant proteins as shown in Figure 4.
Figure 3: Histone H3 trimethyl Lys9 ELISA (H3K9).
The Histone H3 trimethyl Lys9 ELISA was used to assay purified HeLa core histones (250 - 500 ng) made using Active Motif's Histone Purification Mini Kit (Catalog No. 40026) and HeLa acid extracts (5 - 10 µg). The provided Recombinant Histone H3 trimethyl Lys9 protein was assayed from 7.8125 - 500 ng/well as a reference standard curve. Data shown are the results from wells assayed in duplicate. These results are provided for demonstration only.
Figure 4: Specificity of the Histone H3 trimethyl Lys9 ELISA (H3K9).
Recombinant Histone H3, mono-, di- and trimethyl Lys9 proteins were assayed from 15 ng - 1 µg per well using the Histone H3 trimethyl Lys9 ELISA. These results indicate the specificity of the assay. There is extremely low background from histone H3 and little cross-reactivity for mono- or diimethyl Lys9. This means that small, specific changes in trimethyl Lys9 levels can easily be detected with this kit.
The Histone ELISA Method
The Histone Modification ELISA kits are sandwich ELISAs for screening lysine methylation on 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 modification specific rabbit polyclonal antibody detects the lysine methylation, 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 protein made using Active Motif's patented 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.
Contents & Storage
Histone H3 dimethyl Lys9 ELISA Kits include a 96-stripwell Histone H3 Capture plate, a primary antibody specific for Histone H3 dimethyl Lys9, HRP-conjugated anti-rabbit IgG secondary antibody, Assay Dilution Buffer, 20X Wash Buffer, Developing Solution, Stop Solution, Recombinant Histone H3 dimethyl Lys9 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.
Histone H3 trimethyl Lys9 ELISA Kits include a 96-stripwell Histone H3 Capture plate, a primary antibody specific for Histone H3 trimethyl Lys9, HRP-conjugated anti-rabbit IgG secondary antibody, Assay Dilution Buffer, 20X Wash Buffer, Developing Solution, Stop Solution, Recombinant Histone H3 trimethyl Lys9 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.
