Human AFP ELISA Kit

in vitro diagnostic, Hepa AFP-IC is anenzyme linked immunosorbent assay (ELISA) for the quantitative measurement ofalpha-fetoprotein (AFP) immune complexes (AFP-IgM) in serum samples.

alpha-fetoprotein

NM_001134

104150

metal ion binding

1.Reagent And Sample Preparation 1) Reconstitutechromogen solution with 20 mL of distilled water. 2) Reconstitutedilution buffer with 25 mL of distilled water. 3) Reconstitutewashing buffer with 1 L of distilled water. 4) Prepare therequired amount of enzyme-conjugated secondary antibody solution diluting10-fold in reconstituted dilution buffer. 2. AssaySteps 1) Prepareassay reagents as described above. 2) Set up themicrotiter plate with sufficient wells to enable the running of all requiredstandards and samples. 3) Removeexcess microtiter plate strips from the frame and store in the resealablefoil bag with the desiccant provided. 4) Wash themicrotiter plate strips three times with washing buffer (300 uL/well). 5) Dispense100 uL/well ofstandard calibrators (in duplicate) starting from 250 AU/mL and performingin-plate 2-fold serial dilutions to a final concentration of 15, 6 AU/mL. Usedilution buffer as diluent. Also dispense 100 uL/well ofdilution buffer as blank, in duplicate. 6) Dispense100 uL/well ofeight fold (1:8) dilution of samples (in duplicate). Use dilution buffer asdiluent. 7) Incubate1hour at room temperature. 8) Wash sixtimes with washing buffer (300 uL/well). 9) Add 100 uL/well ofdiluted enzyme-conjugated secondary antibody solution. 10) Incubate1h at room temperature. 11) Wash sixtimes with washing buffer (300 uL/well). 12) Preparethe required amount of chromogen-enzyme substrate solution adding 1 uL ofenzyme substrate solution per 3 mL of chromogen solution. Thechromogen-enzyme substrate solution must be used within 24 hours. 13) Apply 150uL/well offreshly prepared chromogen-enzyme substrate solution. Allow color to developfor 20 min. at 37C in the dark and measure OD values of each well using anELISA plate reader set to 405 nm. Plot the standard curve deltaOD valuesas described in the next section: Processing of the results.