The EpiQuik™ In Situ Histone H3K4 Methylation Assay Kit is a convenient package of tools that allows the experimenter to specifically measure histone H3K4 methylation in situ using cultured adherent cells. The kit has the following advantages:
- Quick and efficient procedure, which can be finished within 3 hours
- Innovative colorimetric assay without the need for radioactivity, electrophoresis, or chromatography
- Measurement of in situ histone H3K4 methylation without the need to prepare cell lysates
- Microplate format makes the assay suitable for high throughput analysis of agents that increases or inhibits H3-K4 methylation
- Simple, reliable, and consistent assay conditions
Epigenetic activation or inactivation of genes play a critical role in many important human diseases, especially in cancer. A major mechanism for epigenetic inactivation of the genes is methylation of CpG islands in genome DNA caused by DNA methyltransferases. Histone methyltransferases (HMTs) control or regulate DNA methylation through chromatin-dependent transcription repression or activation. HMTs transfer 1-3 methyl groups from S-adenosyl-Lmethionine to the lysine and arginine residues of histone proteins. SET1, SET7/9, Ash1, ALL-1, MLL, ALR, Trx, and SMYD3 are histone methyltransferases that catalyze methylation of histone H3 at lysine 4 (H3-K4) in mammalian cells. H3-K4 methylation may serve as a global epigenetic mark in euchromatin and mediates activated transcription. Increased global H3-K4 methylation is also found to be involved in some pathological processes such as cancer progress.
Principle & Procedure
The EpiQuik™ In Situ Histone H3K4 Methylation Assay Kit is a whole cell-based detection method for methylated H3-K4. In this assay, adherent cells are cultured in conventional 96-well microplates. After your experimental treatment, cells are fixed and permeabilized. The methylated H3K4 is then detected by an anti-methyl H3-K4 antibody. The ratio or amount of methylated H3K4 can be quantified through HRP conjugated secondary antibody-color development system and is proportional to the intensity of color development.
GA1 (10X Wash Buffer)
GA2 (Permeabilizing Buffer)
GA3 (Blocking Buffer)
GA4 (Antibody Buffer)
GA5 (Capture Antibody, 100 µg/ml)*
GA6 (Detection Antibody, 200 µg/ml)*
GA7 (Developing Solution)
GA8 (Stop Solution)
30% H2O2 Solution
Methylated H3-K4 Control (20 µg/ml)
8-Well Control Strips
* For maximum recovery of the products, centrifuge the original vial prior to opening the cap.