A protocol I have been sent for an ELISA-type experiment uses 0.05% (v/v) Tween-20 in PBS as a detergent to remove weakly bound proteins from a microtiter plate. Since we do not have Tween-20, can this be substituted with Triton X-100 or SDS? If so, in what amount?
Dear Ryan Howard
Yes, you could substitute Tween-20 with either Triton X-100 or SDS as detergent for ELISA plates washing buffer, but its cheaper not poisonous as SDS (Tween as liquid soap).
Pls. find the attached files may help in your research.
http://www.labome.com/method/Detergents-Triton-X-100-Tween-20-and-More.html
https://tools.thermofisher.com/content/sfs/brochures/D19564.pdf
Regards
Prof. Houda Kawas
Although I havent triedto replace Tween 20 by any of them, I would be careful with SDS. I have used SDS to treat antigen (after coating) and some of the antibodies lost reactivity against treated antigen, while others were still reactive. Using SDS for washing could modify your results in some cases.
Besides my own experience I found a study on the subject by Thermo scientific and they say that SDS is too harsh and reduced the specific signals. One of their conclusions is
Detergents with net charges like SDS and DTAB must be avoided because of their disadvantageous interferences with the assay reactants.
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