The EpiQuik™ Circulating Histone H3 Citrullination ELISA Kit (Colorimetric) is a complete set of optimized buffers and reagents specifically designed to quantitatively detect circulating histone H3cit from plasma and serum. This 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 H3cit with no cross-reactivity with unmodified H3 or other modifications at the same arginine site.
- The control is conveniently included for the quantification of H3cit.
- Strip microplate format makes the assay flexible: manual or high throughput.
- Simple, reliable, and consistent assay conditions.
Arginine histone citrullination is one of the many important epigenetic marks, and is essential for the regulation of epigenetic chromatin remodeling and immune cells’ extracellular trap processes. Citrullinated arginine of histone H3 (Arg2, 8, 17) regulates gene expression and is mainly catalyzed by peptidylarginine deiminase 4 (PAD4). Furthermore, citrullination of histones, in particular histone H3, was revealed as a convergence point for diverse inflammatory signals that trigger the neutrophil response to infections. It was reported that citrullinated histone H3 could be a potential serum biomarker for the early diagnosis of septic shock. More immunological diseases such as multiple sclerosis and rheumatoid arthritis seem to be also associated with change of histone H3 citrullination. The circulating histone H3 citrullination can be changed by inhibition or activation of PADs. Therefore, quantitative detection of circulating histone H3 citrullination would provide useful information for better understanding epigenetic regulation of gene activation and silencing, as well as for developing PAD-targeted drugs.
Principle & Procedure
This kit is designed for measuring total H3cit in plasma or serum. In an assay with this kit, the citrullinated histone proteins in the plasma/serum sample are captured on the strip wells coated with anti-citrullinated H3 antibody. The captured citrullinated H3 can then be recognized with a detection antibody followed by a color development reagent. The ratio of H3cit is proportional to the intensity of absorbance. The absolute amount of H3cit 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 FilmDAb (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.