Aug 15, 2020
Binding properties of immunoglobulin-binding protein by double immunodiffusion (Ouchterlony) technique.
- 1University of the West Indies St. Augustine
- University of the West Indies
- angel.vaillant@sta.uwi.edu
Protocol Citation: Angel A Justiz-Vaillant 2020. Binding properties of immunoglobulin-binding protein by double immunodiffusion (Ouchterlony) technique.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjsjkncn
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 15, 2020
Last Modified: August 15, 2020
Protocol Integer ID: 40491
The binding of SpLAG, SpLG and SpAG with animal sera, avian IgY, and purified mammalian IgG are investigated by double immunodiffusion.
Briefly, 1% agarose gels are prepared and wells cut into the gel using a template.
Initially, aliquots of 25 µl each of the immunoglobulin-bacterial protein SpLAG, SpLG or SpAG at a concentration of one (1) µg/µl are applied to the centre well.
The peripheral wells are filled with 25 µl each of IgY (30 µg/µl) or mammalian IgG or sera.
The gels are incubated at RT for 48−72 hours.
After that the gels are examined for precipitin lines.
Human serum or human IgG are included as positive controls.
The positive results are taken as the presence of precipitin line/s and negative results, the absence of precipitin lines.