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Epigenetics & Chromatin

Histone H3 methylated Lys4 ELISA (H3K4)

to quantitate the level of histone H3 lysine 4 methylation

Histone Modification ELISAs provide a simple, sensitive method for detecting changes in specific histone modifications from purified core histones or histones isolated by acid extraction. These kits are sandwich ELISAs that utilize a capture antibody against histone H3 and a primary antibody specifc for the modification of interest. 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 low or high throughput screening. For complete details, click the ELISA Method tab below.

Histone H3 lysine 4 methylation (H3K4) ELISA

Histone H3 lysine 4 methylation ELISA kits provide everything needed for screening the levels of histone H3 monomethyl, dimethyl or trimethyl Lys 4 in human, mouse, rat and other (wide range predicted) systems. Histone H3 lysine 4 methylation (H3K4) has been correlated with euchromatin formation and transcriptional activation, making the methylation state of lysine 4 an interesting marker of transcriptional activity. 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. 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. Click the H3K4 Info tab below for data and more information; kit manuals can be downloaded under the Documents tab.

 
Name Format Cat No. Price  
Histone H3 monomethyl Lys4 ELISA 1 x 96 rxns 53101 $525 Buy Now
Histone H3 dimethyl Lys4 ELISA 1 x 96 rxns 53112 $525 Buy Now
Histone H3 trimethyl Lys4 ELISA 1 x 96 rxns 53113 $525 Buy Now

Histone H3 methylated Lys4 (H3K4)

The methylation of lysine 4 on histone H3 (H3K9) has been correlated with the regulation of gene transcription, making methylated lysine 4 on histone H3 a significant marker in studying the state of transcription activity. Methylation of histone H3 at lysine 4 is linked to euchromatin formation and transcriptional activation.

 

Histone H3 monomethyl Lys4 ELISA

The Histone H3 monomethyl Lys4 ELISA is capable of detecting the level of histone H3 monomethylated at lysine 4 from core histone preparations or histones purified by acid extraction from tissue or cell samples (Figure 1). The Histone H3 monomethyl Lys4 sandwich ELISA assay is also tested for cross-reactivity with other degree-specific recombinant proteins as shown in Figure 2.

 

Histone H3 monomethyl Lys4 ELISA
 
Figure 1: Histone H3 monomethyl Lys4 ELISA (H3K4).

The Histone H3 monomethyl Lys4 ELISA was used to assay purified HeLa core histones (1- µg) made using Active Motif's Histone Purification Mini Kit (Catalog No. 40026) and HeLa acid extracts (10 µg). The provided Recombinant Histone H3 monomethyl Lys4 protein was assayed from 3.9 - 250 ng/well as a reference standard curve. Data shown are the results from wells assayed in duplicate. These results are provided for demonstration only.

 

Specificity of Histone H3 monomethyl Lys4 ELISA
 
Figure 2: Specificity of the Histone H3 monomethyl Lys4 ELISA (H3K4).

Recombinant Histone H3, mono-, di- and trimethyl Lys4 proteins were assayed from 3.9 ng - 250 ng per well using the Histone H3 monomethyl Lys4 ELISA. These results indicate the specificity of the assay. There is extremely low background from histone H3 and little cross-reactivity for di- or trimethyl Lys4. This means that small, specific changes in monomethyl Lys4 levels can easily be detected with this kit.

 

Histone H3 dimethyl Lys4 ELISA

The Histone H3 dimethyl Lys4 ELISA is capable of detecting the level of histone H3 dimethylated at lysine 4 from core histone preparations or histones purified by acid extraction from tissue or cell samples (Figure 3). The Histone H3 dimethyl Lys4 sandwich ELISA assay is also tested for cross-reactivity with other degree-specific recombinant proteins as shown in Figure 4.

 

Histone H3 dimethyl Lys4 ELISA
 
Figure 3: Histone H3 dimethyl Lys4 ELISA (H3K4).

The Histone H3 dimethyl Lys4 ELISA was used to assay purified HeLa core histones (10 µg) made using Active Motif's Histone Purification Mini Kit (Catalog No. 40026) and HeLa acid extracts (10 µg). The provided Recombinant Histone H3 dimethyl Lys4 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.

 

Specificity of Histone H3 dimethyl Lys4 ELISA
 
Figure 4: Specificity of the Histone H3 dimethyl Lys4 ELISA (H3K4).

Recombinant Histone H3, mono-, di- and trimethyl Lys4 proteins were assayed from 1.5 ng - 100 ng per well using the Histone H3 dimethyl Lys4 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 Lys4. This means that small, specific changes in dimethyl Lys4 levels can easily be detected with this kit.

 

Histone H3 trimethyl Lys4 ELISA

The Histone H3 trimethyl Lys4 ELISA is capable of detecting the level of histone H3 trimethylated at lysine 4 from core histone preparations or histones purified by acid extraction from tissue or cell samples (Figure 5). The Histone H3 trimethyl Lys4 sandwich ELISA assay is also tested for cross-reactivity with other degree-specific recombinant proteins as shown in Figure 6.

 

Histone H3 trimethyl Lys4 ELISA
 
Figure 5: Histone H3 trimethyl Lys4 ELISA (H3K94).

The Histone H3 trimethyl Lys4 ELISA was used to assay purified HeLa core histones (1- µg) made using Active Motif's Histone Purification Mini Kit (Catalog No. 40026) and HeLa acid extracts (10 µg). The provided Recombinant Histone H3 trimethyl Lys4 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.

 

Specificity of Histone H3 trimethyl Lys4 ELISA
 
Figure 6: Specificity of the Histone H3 trimethyl Lys4 ELISA (H3K4).

Recombinant Histone H3, mono-, di- and trimethyl Lys9 proteins were assayed from 1.56 - 100 ng per well using the Histone H3 trimethyl Lys4 ELISA. These results indicate the specificity of the assay. There is extremely low background from histone H3 and little cross-reactivity for mono- or dimethyl Lys4. This means that small, specific changes in trimethyl Lys4 levels can easily be detected with this kit.

The Histone Modification ELISA Advantage

Historically, screening for histone modifications from sample preparations have been conducted using immunoblotting and chromatin immunoprecipitation methods, which can be time-consuming, do not allow for high-throughput and provide only semi-quantitative results. Active Motif's Histone Modification ELISAs provide a fast, sensitive assay with the flexibility to screen from 1 to 96 samples in a single experiment.

Included in each Histone Methylation ELISA is a methylated recombinant histone made using Active Motif's patented protein synthesis technology. 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. Each Histone Phosphorylation ELISA contains treated and untreated acid extracts for use as a positive and negative controls.

Why use the Histone Methylation and Histone Phosphorylation ELISAs?

  • Increased sensitivity over immunoblotting methods
  • Results in less than three hours
  • Specific antibody detection ensures low background and no cross-reactivity with other modifications
  • Colorimetric readout enables easy, quantitative analysis with spectrophotometry at 450 nm
  • 96-stripwell format enables both high and low throughput
  • Positive controls included in each kit

The Histone Methylation ELISA Method

The Histone ELISAs to detect site- and degree-specific lysine methylation levels on histone H3 are sandwich ELISAs that utilize a monoclonal histone H3 antibody to capture histone H3 from purified core histones or histones isolated by acid extraction from tissue or cell samples. A polyclonal antibody specific for the modification of interest 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 at 450 nm. The assay is performed in a convenient 96-stripwell plate, which enables either low or high throughput screening.

The Histone Phosphorylation ELISA Method

The Histone ELISAs to detect phosphorylation levels of serine 10 (pSer10) or serine 28 (pSer28) on histone H3 are sandwich ELISAs that utilize a C-terminal histone H3 antibody to capture histone H3 from purified core histones or histones isolated by acid extraction from tissue or cell samples. A monoclonal antibody specific for the phosphorylation site of interest 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 at 450 nm. The assay is performed in a convenient 96-stripwell plate, which enables either low or high throughput screening.

Flow Chart of Histone Modification ELISA Method

Flow chart of the Histone Modification ELISA method.
 
Figure 1: Flow chart of the Histone Modification ELISA Method.

Histone Modification ELISAs utilize a histone H3 antibody to capture histone H3 from samples and a modification specific antibody for detection. A secondary antibody conjugated to horseradish peroxidase and developing solutions provide a colorimetric readout that is quantified by spectrophotometry. (Click image to enlarge.)

Contents & Storage

Histone H3 monomethyl Lys4 ELISA Kits include a 96-stripwell Histone H3 Capture plate, a primary antibody specific for Histone H3 monomethyl Lys4, HRP-conjugated anti-rabbit IgG secondary antibody, Assay Dilution Buffer, 20X Wash Buffer, Developing Solution, Stop Solution, Recombinant Histone H3 monomethyl Lys4 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 dimethyl Lys4 ELISA Kits include a 96-stripwell Histone H3 Capture plate, a primary antibody specific for Histone H3 dimethyl Lys4, HRP-conjugated anti-rabbit IgG secondary antibody, Assay Dilution Buffer, 20X Wash Buffer, Developing Solution, Stop Solution, Recombinant Histone H3 dimethyl Lys4 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 Lys4 ELISA Kits include a 96-stripwell Histone H3 Capture plate, a primary antibody specific for Histone H3 trimethyl Lys4, HRP-conjugated anti-rabbit IgG secondary antibody, Assay Dilution Buffer, 20X Wash Buffer, Developing Solution, Stop Solution, Recombinant Histone H3 trimethyl Lys4 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.