PADI4 Fluorogenic Assay Kit
The PADI4 Fluorogenic Assay Kit is a fluorogenic assay designed to measure the enzymatic activity of PADI4 (protein-arginine deiminase type-4) for screening and profiling applications. This assay uses o-phthalaldehyde and DTT (dithiothreitol) as detection reagents. The assay kit comes in a convenient 96-well format, with enough purified recombinant PADI4, BAEE (Nα-Benzoyl-L-arginine ethyl ester hydrochloride), EDTA (Ethylenediaminetetraacetic acid), o-phthalaldehyde, and assay buffer for 100 reactions.
Figure 1: Assay Principle. Peptidyl-arginine is converted into peptidyl-citrulline by PADI4, releasing NH4+. NH4+ reacts with DTT-reduced o-phthalaldehyde. O-phthalaldehyde can then fluoresce at 470 nm, and the signal is proportional to the activity of PADI4.
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- Ethanol
- Fluorescence plate reader capable of measurement at λex413/λem470 nm
- Adjustable micropipettor and sterile tips
- 37°C incubator
| Catalog # | Name | Amount | Storage |
| 50154 | PADI4, FLAG-Tag* | 40 µg | -80°C |
| PADI Assay Buffer | 10 ml | -20°C | |
| 0.5 M EDTA | 1.5 ml | -20°C | |
| 100 mM BAEE | 500 µl | -80°C | |
| 100 mM O-phthalaldehyde (OPT) | 1.5 ml | +4°C | |
| 0.5 M DTT | 200 μl | -80°C | |
| 100 mM Phosphate Buffer, pH 7.8 | 20 ml | Room Temp | |
| 79685 | Black 96-well plate | 2 | Room Temp |
*The concentration of the protein is lot-specific and will be indicated on the tube
PADI4, also known as peptidylarginine deiminase 4, is a hydrolase that catalyzes the conversion of peptidylarginine to citrulline, a process known as citrullination. PADI4 plays a role in several cellular functions, including gene expression, cell differentiation, and apoptosis. It is involved in immune responses and inflammation through granulocyte and macrophage development and thus contributes to diseases such as RA (rheumatoid arthritis). Dysfunction of PADI4 has also been linked to multiple sclerosis (MS), Alzheimer’s disease (AD) and cancer. It contributes to cancer by several possible mechanisms such as epithelial-to-mesenchymal transition (EMT), apoptosis, and formation of neutrophil extracellular traps (NETs). A better understanding of the contribution of PADI4 to cancer development and inflammation, combined with development of inhibitors, will result in new therapeutic avenues.
Wu X., et al., 2023 Anticancer Agents Med Chem 23(3): 256-265
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