Bovine NEFA (Non-ester fatty acid) ELISA Kit

From Antibodies.com, this competitive Enzyme-Linked Immunosorbent Assay (cELISA) is designed for the in vitro quantitative determination of bovine non-esterified fatty acids (NEFA) in serum, plasma, tissue homogenates, and other biological fluids. The 2-hour (120 minute) colorimetric assay uses a pre-coated 96-well microplate arranged as 12 × 8 well strips, with absorbance read at 450 nm.

During the run, NEFA in standards or samples competes with immobilised antigen for binding to a biotinylated anti-NEFA antibody. After washing, HRP-Streptavidin conjugate and TMB substrate are added, producing a blue product that turns yellow on addition of the acidic stop solution. The intensity of the yellow colour is inversely proportional to the NEFA concentration, which is read against the standard curve. Reported sensitivity is 3.75 nmol/mL and the working range is 6.25–400 nmol/mL. The kit is reported to be specific for non-esterified fatty acids with no cross-reactivity with other analogs or variants.

Validation summaries give neat-sample recoveries (average) of 100% in serum (range 95–103%), 92% in EDTA plasma (87–105%) and 90% in heparin plasma (87–93%), with dilution linearity at 1:2, 1:4 and 1:8 of 85–104%, 92–100% and 87–96% in serum, and comparable ranges for EDTA and heparin plasma. Supplied components include a pre-coated microplate, lyophilised standard, sample dilution buffer, biotinylated detection antibody, antibody dilution buffer, HRP-Streptavidin conjugate with its dilution buffer, TMB substrate, stop solution, 25× wash buffer, plate sealers, and a foil pouch. Kit components should be stored at +4°C and the kit is shipped at +4°C. The product is for research use only and is not intended for diagnostic or therapeutic use.