The EpiQuik™ Circulating Acetyl Histone H3K56 ELISA Kit (Colorimetric) is a convenient package of tools that allows the experimenter to specifically measure circulating acetyl histone H356 (H3K56ac) from biological fluid samples such as plasma and serum from human, mouse or rat. The kit has the following advantages:
- Quick and efficient procedure, which can be finished within 2.5 hours.
- High sensitivity and specificity. The detection limit is as low as 0.5 ng/well with dynamic range of 1-20 ng/well within the indicated amount range of the plasma/serum. Only recognizes H3K56ac with no cross-reactivity with unmodified H3 or other modifications at the same lysine site.
- The control is conveniently included for the quantification of H3K56ac.
- Strip microplate format makes the assay flexible: manual or high throughput.
- Simple, reliable, and consistent assay conditions.
Epigenetic activation or inactivation of genes plays a critical role in many important human diseases, especially in cancer. A major mechanism for epigenetic gene inactivation is methylation of CpG islands in genomic DNA caused by DNA methyltransferases. Histone acetyltransferases (HATs) and deacetylases (HDACs) control or regulate DNA methylation through chromatin-dependent transcription repression or activation. HATs transfer acetyl groups from acetyl CoA to the lysine residues of histone proteins. P300 and CBP are the major HATs that catalyzes acetylation of histone H3 at lysine 56 (H3K56) in mammalian cells. HDAC1 and SIRT 1/3/6 are the histone deacetylases that deacetylate H3K56. H3K56ac has been viewed as a signature mark of transcriptionally activate genes, which is placed exclusively in the 5’- region downstream of the promoter. The H3K56ac can also be changed by inhibition or activation of HATs or HDAC/SIRTs. Circulating histone H3K56ac in plasma or serum has been observed and demonstrated as the marker for many different diseases or pathological changes such as cancer progression. Therefore, detection of circulating H3K56ac would provide useful information for a better understanding of epigenetic regulation of gene activation and silencing, histone modification-associated pathological processes, screening of disease-related biomarkers, as well as for developing histone modification-targeted drugs.
Principle & Procedure
The EpiQuik™ Circulating Acetyl Histone H3K56 ELISA Kit (Colorimetric) is designed for measuring total H3K56ac in plasma or serum. In an assay with this kit, the Histone H3 proteins acetylated at K56 in the plasma/serum sample are captured on the strip wells coated with anti-H3K56ac antibody. The captured H3K56ac proteins can be then recognized with detection antibody followed by a color development reagent. The ratio of H3K56ac is proportional to the intensity of absorbance. The absolute amount of H3K56ac can be quantitated by comparing to the standard control.
Input materials should be plasma or serum. The amount of plasma or serum for each assay can be 10 to 40 µl with an optimal amount of 30 µl.
WB (10X Wash Buffer)
HAB (Histone Assay Buffer)
DS (Developer Solution)
SS (Stop Solution)
8-Well Assay Strips (With Frame)
Control Assay Strips (With Frame)#
Adhesive Covering Film
DAb (Detection Antibody, 1000X)*
Standard Control (100 µg/ml)
*Spin the solution down to the bottom prior to use.
#Control Assay Strips are green trimmed for distinguishing from 8-well Assay Strips (for samples). The Control Assay Strips are only for control use and should not be used for sample assay.