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Roche protease inhibitor 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 protease inhibitor might interfere in assays involving particular bead products. 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 at 1X
Catalog NumberProduct% of inhibition at max conc
AL125Streptavidin AlphaLISA Acceptor Beads0
AL126Protein L AlphaLISA Acceptor Beads31
AL127Anti-FITC AlphaLISA Acceptor Beads51
AL128 (biotin)Anti-6xHis AlphaLISA Acceptor Beads0
AL128 (GSH)Anti-6XHis AlphaLISA Acceptor Beads85
AL129Anti-V5 AlphaLISA Acceptor Beads0
AL130Anti-mouse IgM AlphaLISA Acceptor Beads24
AL131Anti-chicken IgY AlphaLISA Acceptor Beads0
AL132Anti-sheep IgG AlphaLISA Acceptor Beads45
AL133Anti-GFP AlphaLISA Acceptor Beads51
AL134Anti-MBP AlphaLISA Acceptor Beads37
AL136Strep-Tactin AlphaLISA Acceptor Beads80
AS102Protein A Alpha Donor Beads0
AS103Anti-FLAG Alpha Donor Beads81
AS104Anti-mouse IgG Alpha Donor Beads32
AS105Anti-rabbit IgG Alpha Donor Beads29
AS106Strep-Tactin Alpha Donor Beads0
AL900AlphaLISA TruHits kit 2 ug/mL0
AL900AlphaLISA TruHits kit 20 ug/mLnd

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 protease inhibitor using anti-6X His AlphaLISA Acceptor beads (with glutathione Donor beads) showed 85% signal inhibition when 1X protease inhibitor was used.

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.