PADI4 Fluorogenic Assay Kit

• Background:

  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.

• Description:

  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.Need us to run inhibitor screens or profile your compounds against PADI4? Check out ourMetabolic Enzyme Screening Services.

• UniProt:

  Q9UM07

• Applications:

  Study enzyme kinetics and screen small molecule inhibitors for drug discovery and high throughput screening (HTS) applications.

• Format:

  Catalog #NameAmountStorage50154PADI4, FLAG-Tag*40 µg-80°CPADI Assay Buffer10 ml-20°C0.5 M EDTA1.5 ml-20°C100 mM BAEE500 µl-80°C100 mM O-phthalaldehyde (OPT) 1.5 ml+4°C0.5 M DTT200 μl-80°C100 mM Phosphate Buffer, pH 7.820 mlRoom Temp79685Black 96-well plate2Room Temp*The concentration of the protein is lot-specific and will be indicated on the tube

• Shipping Conditions:

  -80°C

• Storage Conditions:

  This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

• Notes:

  Troubleshooting GuideVisitbpsbioscience.com/assay-kits-faqfor detailed troubleshooting instructions. For all further questions, please email[email protected]

• Contraindications:

  The final concentration of DMSO in the assay should not exceed 1%.