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Glycerol 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 glycerol might interfere in assays involving particular bead products, and at what glycerol 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 30%
Catalog NumberProductNo effect (10%)50% loss% of inhibition at max conc
AL125Streptavidin AlphaLISA Acceptor Beads0.23.991
AL126Protein L AlphaLISA Acceptor Beads2.58.480
AL127Anti-FITC AlphaLISA Acceptor Beads2.48.185
AL128 (biotin)Anti-6xHis AlphaLISA Acceptor Beads0.56.487
AL128 (GSH)Anti-6XHis AlphaLISA Acceptor Beads0.88.687
AL129Anti-V5 AlphaLISA Acceptor Beads1.57.087
AL130Anti-mouse IgM AlphaLISA Acceptor Beads1.23.294
AL131Anti-chicken IgY AlphaLISA Acceptor Beads1.04.490
AL132Anti-sheep IgG AlphaLISA Acceptor Beads1.17.491
AL133Anti-GFP AlphaLISA Acceptor Beads0.32.996
AL134Anti-MBP AlphaLISA Acceptor Beads1.35.886
AL136Strep-Tactin AlphaLISA Acceptor Beads0.211.780
AS102Protein A Alpha Donor Beads0.32.597
AS103Anti-FLAG Alpha Donor Beads0.98.388
AS104Anti-mouse IgG Alpha Donor Beads0.12.194
AS105Anti-rabbit IgG Alpha Donor Beads0.23.991
AS106Strep-Tactin Alpha Donor Beads21.527.365
AL900AlphaLISA TruHits kit 2 ug/mL12.024.085
AL900AlphaLISA TruHits kit 20 ug/mL2.014.084

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 glycerol using anti-6X His AlphaLISA Acceptor beads (with glutathione Donor beads) showed little effect on the assay signal at final glycerol concentrations up to ~0.8%. Glycerol did show a half-maximal inhibition of signal when used at a final concentration of 8.6%. When 30% glycerol was included in the assay, 87% 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.