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IBMX 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 IBMX might interfere in assays involving particular bead products, and at what IBMX 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 0.0025M
Catalog NumberProductNo effect (10%)50% loss% of inhibition at max conc
AL125Streptavidin AlphaLISA Acceptor Beads1.0E-06>0.002537
AL126Protein L AlphaLISA Acceptor Beads0.0E+00>0.002535
AL127Anti-FITC AlphaLISA Acceptor Beads1.1E-03>0.002542
AL128 (biotin)Anti-6xHis AlphaLISA Acceptor Beads>0.0025>0.00250
AL128 (GSH)Anti-6XHis AlphaLISA Acceptor Beads>0.0025>0.00250
AL129Anti-V5 AlphaLISA Acceptor Beads5.3E-061.5E-0459
AL130Anti-mouse IgM AlphaLISA Acceptor Beads1.2E-054.2E-0553
AL131Anti-chicken IgY AlphaLISA Acceptor Beads1.4E-03>0.002539
AL132Anti-sheep IgG AlphaLISA Acceptor Beads1.0E-03>0.002546
AL133Anti-GFP AlphaLISA Acceptor Beads2.5E-043.3E-0459
AL134Anti-MBP AlphaLISA Acceptor Beads2.4E-04>0.002525
AL136Strep-Tactin AlphaLISA Acceptor Beads>0.0025>0.00250
AS102Protein A Alpha Donor Beads6.4E-04>0.002548
AS103Anti-FLAG Alpha Donor Beads>0.0025>0.00250
AS104Anti-mouse IgG Alpha Donor Beads1.0E-03>0.002540
AS105Anti-rabbit IgG Alpha Donor Beads1.0E-06>0.002537
AS106Strep-Tactin Alpha Donor Beads>0.0025>0.00250
AL900AlphaLISA TruHits kit 2 ug/mL>0.0025>0.00250
AL900AlphaLISA TruHits kit 20 ug/mL2.2E-03>0.002524

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 IBMX using anti-6X His AlphaLISA Acceptor beads (with glutathione Donor beads) showed little effect on the assay signal at final IBMX concentrations up to 2.5 mM.

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.