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Urea and bead interference

The table below shows results from a compound interference study. We tested a panel of 24 compounds in a QC assay using our AlphaLISA® toolbox bead products. The information below can be used for guidance as to whether urea might interfere in assays involving particular bead products, and at what urea concentration interference might occur. These data are derived from single experiments and should serve more as a guide rather than a precise value, as it is unlikely you will be using the exact same assay design as used to generate the data below. Interference concentrations may vary depending on the assay components. It is possible your assay will tolerate higher or lower concentration than what is shown.



Tested up to 0.1M
Catalog NumberProductNo effect (10%)50% loss% of inhibition at max conc
AL125Streptavidin AlphaLISA Acceptor Beads>0.1>0.10
AL126Protein L AlphaLISA Acceptor Beads>0.1>0.10
AL127Anti-FITC AlphaLISA Acceptor Beads3.8E-02>0.121
AL128 (biotin)Anti-6xHis AlphaLISA Acceptor Beads9.3E-02>0.113
AL128 (GSH)Anti-6XHis AlphaLISA Acceptor Beads2.0E-02>0.119
AL129Anti-V5 AlphaLISA Acceptor Beads>0.1>0.10
AL130Anti-mouse IgM AlphaLISA Acceptor Beads>0.1>0.10
AL131Anti-chicken IgY AlphaLISA Acceptor Beads>0.1>0.111
AL132Anti-sheep IgG AlphaLISA Acceptor Beads>0.1>0.10
AL133Anti-GFP AlphaLISA Acceptor Beads>0.1>0.10
AL134Anti-MBP AlphaLISA Acceptor Beads>0.1>0.10
AL136Strep-Tactin AlphaLISA Acceptor Beads>0.1>0.10
AS102Protein A Alpha Donor Beads>0.1>0.10
AS103Anti-FLAG Alpha Donor Beads>0.1>0.10
AS104Anti-mouse IgG Alpha Donor Beads>0.1>0.10
AS105Anti-rabbit IgG Alpha Donor Beads>0.1>0.10
AS106Strep-Tactin Alpha Donor Beads>0.1>0.10
AL900AlphaLISA TruHits kit 2 ug/mL>0.1>0.10
AL900AlphaLISA TruHits kit 20 ug/mL>0.1>0.10

Refer to the figure below for more detail as to what the values in the table above indicate. Please note that the figure is provided as a general example to explain how the values in our interference tables were determined. The table above indicates that an assay testing the effect of urea using anti-6X His AlphaLISA Acceptor beads (with glutathione Donor beads) showed little effect on the assay signal at final urea concentrations up to 20 mM. When 0.1 M urea was included in the assay, 19% signal inhibition was observed.

interferencetesting16_ASK.jpg
Figure: Key for how the values in our interference tables are derived. The “no effect” concentration was obtained by extrapolating the MAX - 10% MAX counts. Refer to the table above for actual interference values for each toolbox bead product.