Human KL4/Krueppel-like factor 14 ELISA Kit

Assay Genie provides a sandwich ELISA kit built on a double-antibody configuration for the quantitative measurement of human KL4/Krueppel-like factor 14. The kit targets human samples, including serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples, and is read colorimetrically. Per the manufacturer datasheet, the assay has a stated sensitivity of 0.094 nmol/L and a working range of 0.156–10 pmol/mL.

Linearity performance reported by the manufacturer covers dilutions of 1:2, 1:4, and 1:8. For serum (n = 5) the ranges are 88–101%, 94–101%, and 88–105%; for EDTA plasma (n = 5) 89–97%, 84–101%, and 84–98%; and for heparin plasma (n = 5) 80–97%, 80–97%, and 84–100%. Recovery data for the same three matrices are given as: serum (n = 5) 17–83% with an average of 48%, EDTA plasma (n = 5) 62–102% with an average of 76%, and heparin plasma (n = 5) 6–70% with an average of 44%. The protocol follows standard sandwich ELISA steps, beginning with reagent equilibration and loading of standards, samples, and a zero control onto the pre-coated plate, followed by primary incubation at 37°C for 90 minutes, biotin-labeled detection antibody incubation at 37°C for 60 minutes, HRP-Streptavidin (SABC) incubation at 37°C for 30 minutes, TMB color development in the dark for 10–20 minutes, and absorbance reading at 450 nm after stop solution.

Sample preparation guidance includes allowing blood to clot and centrifuging serum at 1000 × g for 20 minutes, collecting plasma with anticoagulant tubes and centrifuging at 1000 × g for 15 minutes at 2–8°C, homogenising tissue in PBS with protease inhibitors, centrifuging cell culture supernatant at 2500 rpm for 5 minutes, and lysing cells with buffer containing protease inhibitors. Components supplied for the assay format include a dismountable ELISA microplate, lyophilized standard, biotin-labeled antibody concentrate (100X), HRP-Streptavidin conjugate (SABC, 100X), TMB substrate, sample dilution buffer, antibody dilution buffer, SABC dilution buffer, stop solution, 25X wash buffer, plate sealers, and a technical manual.