Infliximab Kit ELISA

• Before performing the assay, samples and assay kit should be brought to room temperature (about 30 minutes beforehand) and ensure the homogeneity of the solution.
• All Standards should be run with each series of unknown samples.
• Standards should be subject to the same manipulations and incubation times as the samples being tested.
• All steps of the test should be completed without interruption. 6
• Use new disposable plastic pipette tips for each reagent, standard or specimen in order to avoid cross contamination.

0.2 - 6 µg/mL (in case of the recommended sample dilution factor)
10 ng/ml (sensitivity in the well)

Just the wash buffer has to be prepared by diluting the stock solution 1:20. All other reagents are ready to use.

dilute the samples 1:20 with assay buffer

Pipette 100 µL of assay buffer into each of the wells.
Pipette 20 µL of each standard and diluted samples.
Incubate 30 min at room temperature (RT).
Aspirate or decant the incubation solution.
Wash the plate 3 x 300 μL of diluted wash buffer per well.
Pipette 100 μL of enzyme conjugate (HRP-anti human IgG mAb).
Shake plate carefully. Incubate 30 min at RT.
Wash the plate 3 x 300 μL of diluted wash buffer per well.
Pipette 100 µL TMB substrate solution into each well.
Incubate 15 min at RT.
Add 100 µL of stop solution.
Measure optical density (OD) at 450 nm.

The results are read from a standard curve.