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Anti-Sheep IgG AlphaLISA Acceptor 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 what buffer components might interfere with AlphaLISA assays using Anti-Sheep IgG AlphaLISA Acceptor beads, and at what concentrations 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.



No effect
10%
50% lossTested up to% of inhibition at max conc
Glycerol%1.17.430%91
DMSO%0.82.310%80
Gelatin%>1>11%0
CHAPS%0.040.23%80
Deoxycholate%0.010.053%92
SDS%0.10.13%98
Triton X-100%>3>33%0
Tween 20%>3>33%0
TRIS (pH 8.0)M1.7E-012.2E-010.5M57
2-MercaptoethanolM7.6E-035.9E-020.1M58
DTTM2.4E-044.3E-020.1M59
EDTAM2.1E-02>0.10.1M28
EGTAM>0.1>0.10.1M0
IBMXM1.0E-03>0.00250.0025M46
ImidazoleM9.3E-045.3E-030.1M99
Mg2+ (MgCl2)M1.0E-02>0.10.1M26
NitroprussideM4.4E-034.5E-020.1M69
UreaM>0.1>0.10.1M0
VanadateM4.9E-032.3E-020.1M74
CitrateM2.5E-033.1E-030.1M100
Adenosine (ATP)M>0.01>0.010.01M0
GlycineM>0.01>0.010.01M0
NaFM>0.01>0.010.01M0
S-adenosylmethionineM>0.001>0.0010.001M0
Complete Protease InhibitorXnot determinednot determined1X45

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 different chemicals using Anti-Sheep IgG AlphaLISA Acceptor beads (with Streptavidin Donor beads) showed little effect on the assay signal at final Triton X-100 concentrations up to 3%. Using 7.4% glycerol resulted in a 50% loss of signal.

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.