Aug 28, 2020
Enzyme linked immunosorbent assay (ELISA) for determining the serum concentration of IL-33 in humans.
- 1University of the West Indies St. Augustine
- University of the West Indies
- angel.vaillant@sta.uwi.edu
Protocol Citation: Angel A Justiz-Vaillant 2020. Enzyme linked immunosorbent assay (ELISA) for determining the serum concentration of IL-33 in humans.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjy9kpz6
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 19, 2020
Last Modified: August 28, 2020
Protocol Integer ID: 40705
Ninety-six well ELISA plates are coated with monoclonal anti-human antibodies to interleukin-33 (IL-33).
Patient serum samples are added to the plates.
The plate is incubate x 1.30 hour at RT.
The plate is washed 4 times with PBS-tween buffer.
The wells are incubated with a biotin conjugated anti-human IL-33 for 1.30 hour at RT.
The plate is washed again as above.
Add to the plate a peroxidase-labeled streptavidin conjugate and incubate it for 1 hour at RT.
After a further washing procedure a substrate solution reactive is added and allowed to produced a colored reaction in positive controls.
The level of IL-33 in the sample is proportional to the colored product developed.
The addition of 3M H2SO4 stops the reaction.
The absorbance is measured at 450 nm.
The IL-33 concentration can be calculated by generating an standard curve using lineal regression.