Description

DYKDDDDK tag has a small unique protein sequence that is commonly fused to N- or C-terminus of a protein for expression. Expression and detection of DYKDDDDK-tag proteins are frequently performed in many academic and industry laboratories because this tag is generally more hydrophilic than other common tags and therefore it is less likely to denature or inactivate proteins to which it is attached. In addition, DYKDDDDK-tag fusion proteins in E. coli or mammalian cell lysate can be easily purified by affinity chromatography using specific resins. Typically DYKDDDDK-tag proteins are detected by SDS-PAGE or western blots but these approaches are generally more laborious, time-consuming and less sensitive. BioVision’s DYKDDDDK-Tag Protein ELISA Kit is based on the competitive ELISA principle and it is an easy, fast and sensitive method to detect the expressed DYKDDDDK-tag proteins. This detection kit offers ready-to-use reagents, and can detect as low as 100 ng of DYKDDDDK-tag protein in approximately 1.5 hrs.

Datasheet

FAQ

Can I use cell culture supernatant as a sample with this ELISA kit?
Yes, you can use cell culture supernatant as a sample.

Could you please explain the principle clearly? Also, does the size of my fusion protein matters in the detection process?
1. E4700 is a competitive ELISA kit, where the FLAG peptide is coated to the 96 well plates and the antibody (against Flag protein) that is provided with the kit will bind to the FLAG-tagged protein in the sample and will be removed during the wash steps. The remaining antibody (unbound) will bind to the wells. This means that the higher the concentration of your FLAG protein, less unbound antibody will bind to the wells, leading to lower OD readings and vice versa. Basically, the competition is between the bound and unbound antibody.
2. Regarding your question whether the size of the fusion protein matters, it is not the size, but it is the amount of the fusion protein that matters. The primary antibody is going to be the limiting factor and the primary antibody included in the kit is enough and not going to be saturated.

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