Human Peptidyl Arginine Deiminase Type IV ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is designed for the in vitro quantitative determination of human PADI4/PAD4 in serum, plasma, tissue homogenates or other biological fluids. The assay employs a pre-coated 96-well microtiter plate to which an antibody specific for PADI4/PAD4 is immobilised. Standards and test samples are added to the wells so that PADI4/PAD4 present is captured, followed by incubation with a Biotinylated Anti-PADI4/PAD4 Antibody and then an HRP-Avidin conjugate. TMB substrate is used to visualise the reaction, producing a blue colour that changes upon addition of acidic stop solution; the resulting yellow intensity is proportional to the amount of PADI4/PAD4 captured, and the concentration is calculated from the O.D. absorbance read at 450 nm against a standard curve using a colorimetric microplate reader.

Reported performance figures include a sensitivity of <0.111 ng/ml and an assay range of 0.312–20 ng/ml, with a total assay time of 4h 30m. Spike-and-recovery data are documented for serum, EDTA plasma and heparin plasma: serum average recovery is 91% (range 80%–102%, n=5), EDTA plasma average is 90% (range 81%–100%, n=5), and heparin plasma average is 84% (range 80%–89%, n=5). Linearity upon dilution from 1:2 through 1:16 is also reported across these same sample matrices (S1).

Kit components are supplied for storage at +4°C or −20°C as instructed, and the kit is intended for use with serum, plasma and tissue homogenates (S1).