Rat Beta-defensin 4/Defb4 ELISA Kit

Assay Genie provides a sandwich ELISA kit configured for the quantitative detection of Beta-defensin 4 (DEFB4) in rat-derived samples. The assay uses a double-antibody sandwich format with colorimetric readout and is intended for research into innate immunity, antimicrobial defense and inflammatory conditions in rats, where DEFB4 acts as an antimicrobial peptide of the innate immune system.

Validated sample types include serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. The kit reports a sensitivity of 0.188 ng/mL and an assay range of 0.313–20 ng/mL. Reported linearity across 1:2, 1:4 and 1:8 dilutions spans 87–103%, 85–105% and 92–105% for serum; 82–98%, 85–99% and 82–100% for EDTA plasma; and 82–98%, 82–98% and 85–96% for heparin plasma. Recovery ranges are 90–102% for serum (average 96%), 89–100% for EDTA plasma (average 94%), and 86–103% for heparin plasma (average 91%). Components should be stored at 2–8°C for up to 12 months, protected from direct light where indicated.

The protocol involves equilibrating reagents and TMB substrate to room temperature, loading standards, samples and blanks onto the pre-coated plate, incubating at 37°C for 90 minutes, adding a biotin-labeled detection antibody with incubation at 37°C for 60 minutes, binding HRP-Streptavidin (SABC) at 37°C for 30 minutes, developing color with TMB substrate for 10–20 minutes in the dark, and reading absorbance at 450 nm after adding the stop solution.