Human Forkhead Box Protein P3 ELISA Kit

From Antibodies.com's Assays and Kits range, this sandwich Enzyme-Linked Immunosorbent Assay is designed for the in vitro quantitative determination of human FOXP3 in tissue homogenates, cell lysates, cell culture supernates or other biological fluids. The kit employs the sandwich enzyme immunoassay technique using a microtiter plate pre-coated with an antibody specific for FOXP3. Standards and test samples are added to the wells, where FOXP3 is captured by the immobilized antibody. After washing, a Biotinylated Anti-FOXP3 Antibody binds the captured analyte, and an HRP-Avidin conjugate is added. TMB substrate solution is then used to visualize the enzymatic reaction, producing a blue-coloured product that changes to yellow upon addition of the acidic stop solution. The density of yellow is proportional to the amount of FOXP3 in each well, and the concentration is calculated by reading the O.D. absorbance at 450 nm in a microplate reader against a standard curve.

The assay is read on a colorimetric microplate reader and has a reported product range of 0.312–20 ng/ml with a sensitivity of <0.119 ng/ml. Total assay time is approximately 4 hours 30 minutes. In validation work, recovery in serum (n=5) ranged from 80% – 102% with an average of 91%, in EDTA plasma (n=5) from 81% – 100% averaging 90%, and in heparin plasma (n=5) from 80% – 89% averaging 84%. Linearity upon sample dilution was reported across 1:2, 1:4, 1:8 and 1:16 dilutions for serum, EDTA plasma and heparin plasma. Kit components should be stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.