Histone H3 acetylated Lys27 (H3K27)
The acetylation of lysine 27 on histone H3 (H3K27ac) has been correlated with active enhancers and promoters, making acetylated lysine 27 on histone H3 a significant marker in studying the state of transcription activity.
Histone H3 acetyl Lys27 ELISA (H3K27ac)
The Histone H3 acetyl Lys27 ELISA is capable of detecting the level of histone H3 acetylated at lysine 27 from core histone preparations or histones purified by acid extraction from tissue or cell samples (Figure 1). The Histone H3 acetyl Lys27 sandwich ELISA assay is also tested for cross-reactivity with other site and degree-specific recombinant proteins as shown in Figure 2.
Figure 1: Histone H3 acetyl Lys27 ELISA (H3K27ac).
The Histone H3 acetyl Lys27 ELISA was used to assay HeLa acid extracts (5 µg) that were either untreated (–) or treated with sodium butyrate (+), a known HDAC inhibitor. The provided Recombinant Histone H3 acetyl Lys27 protein was assayed from 0.67 - 3.75 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 acetyl Lys27 ELISA (H3K27ac).
To verify the specificity of the Histone H3 acetyl Lys 27 ELISA, 100 ng of recombinant histone H3, mono-, di- and trimethyl Lys9 and Lys 27, and acetyl Lys4, Lys9, Lys14, Lys18 and Lys23 proteins were assayed per well. These results show the assay specifically recognizes H3K27ac. There is extremely low background from histone H3 and no cross-reactivity with the other methylated or acetylated histone modifications. This means that small, specific changes in acetyl Lys9 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 Acetylation ELISA is an acetylated recombinant histone protein made using Active Motif's patented protein synthesis technology. Likewise, included in each Histone Methylation ELISA is a methylated recombinant histone protein made using Active Motif's patented protein synthesis technology. These included proteins can be used to build a reference standard curve to quantitate the amount of either specifically acetylated or methylated histone H3 (respectively) in your samples. Click on our link for more information on our patented protein synthesis technologies and to get a complete list of our Recombinant Histones and Modified Histones. 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, Histone Acetylation 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 Acetylation ELISA Method
The Histone ELISAs to detect acetylation 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 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 pero