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we are interested in the quantification of TNF and we are working with
the affibody molecule (Catalog number ab 31908).
In particular we would like to develope an ELISA sandwich assay using
the affibody to modify the plates. We found in the data sheet that a
treatment with some reducing agents (such as DDT) before use it is
suggested in order to eliminate possible dimers formation. Is this
procedure strictly reccomanded even for an ELISA assay?
In this case can you suggest us the best protocol to follow?
Thank you for you kind help.
Asked on Apr 05 2012
Thank you for contacting us.
The sELISA application with ab31908 is based on coating of affibody molecules to the flat-bottomed plate. So we recommend reducing conditions even when using in ELISA.
Recommended reducing condition is 20 mM DTT at a pH above 7.5 and incubation at room temperature for 2 hours. Remove excess DTT by passage through a desalting column, not by dialysis.
Please follow the protocol as given in the protocol section of the datasheet.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Answered on Apr 05 2012