Human Interferon Regulatory Factor 5 ELISA Kit

From Antibodies.com comes a sandwich Enzyme-Linked Immunosorbent Assay (ELISA) intended for the in vitro quantitative measurement of human Interferon Regulatory Factor 5 (IRF5) in serum, plasma, tissue homogenates, and other biological fluids. The assay uses a pre-coated 96-well microplate with colorimetric detection, and absorbance is read at 450 nm on a standard microplate reader. Reported sensitivity is <0.051 ng/ml and the assay range is 0.156–10 ng/ml, with a total run time of 4 hours 30 minutes.

The procedure follows a sandwich immunoassay format: standards and samples are added to wells pre-coated with an antibody specific for IRF5, and the target in each sample is captured during incubation. After washing, a biotinylated anti-IRF5 detection antibody is added, followed by an HRP-avidin conjugate. TMB substrate is then introduced and catalysed by HRP to produce a blue product that turns yellow after addition of stop solution; the absorbance at 450 nm is proportional to IRF5 concentration and is quantified against the standard curve.

Manufacturer-supplied validation includes spike-and-recovery testing with n=5 per matrix: serum averaged 91% (range 80–102%), EDTA plasma averaged 90% (range 81–100%), and heparin plasma averaged 84% (range 80–89%). Dilution linearity across 1:2, 1:4, 1:8, and 1:16 dilutions is also reported for each sample type. Kit components include the pre-coated microplate, lyophilized standard, detection solutions A and B, wash buffer, sample dilution buffer, TMB substrate, stop solution, and adhesive plate sealers, with storage at +4°C or –20°C as instructed.