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Citrate and bead interference

Overview

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 citrate might interfere in assays involving particular bead products, and at what citrate 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 Number Product No effect (10%) 50% loss % of inhibition at max conc
AL125 Streptavidin AlphaLISA Acceptor Beads 1.7E-03 3.9E-03 100
AL126 Protein L AlphaLISA Acceptor Beads 3.0E-03 3.6E-03 100
AL127 Anti-FITC AlphaLISA Acceptor Beads 1.3E-03 2.1E-03 99
AL128(biotin) Anti-6xHis AlphaLISA Acceptor Beads 7.5E-04 8.6E-04 100
AL128(GSH) Anti-6XHis AlphaLISA Acceptor Beads 7.7E-04 9.4E-04 100
AL129 Anti-V5 AlphaLISA Acceptor Beads 2.8E-06 8.9E-05 98
AL130 Anti-mouse IgM AlphaLISA Acceptor Beads 9.5E-04 1.2E-03 100
AL131 Anti-chicken IgY AlphaLISA Acceptor Beads 2.3E-03 2.8E-03 99
AL132 Anti-sheep IgG AlphaLISA Acceptor Beads 2.5E-03 3.1E-03 100
AL133 Anti-GFP AlphaLISA Acceptor Beads 2.1E-03 2.2E-03 100
AL134 Anti-MBP AlphaLISA Acceptor Beads 2.4E-03 2.7E-03 100
AL136 Strep-Tactin AlphaLISA Acceptor Beads 1.4E-03 2.6E-03 100
AS102 Protein A Alpha Donor Beads 7.6E-05 4.9E-04 100
AS103 Anti-FLAG Alpha Donor Beads 2.3E-03 2.5E-03 100
AS104
Anti-mouse IgG Alpha Donor Beads
1.6E-03 2.5E-03 100
AS105 Anti-rabbit IgG Alpha Donor Beads 1.7E-03 3.9E-03 100
AS106 Strep-Tactin Alpha Donor Beads 9.4E-04 1.1E-03 100
AL900  AlphaLISA TruHits kit 2 ug/mL 3.1E-02 >0.1 49
AL900  AlphaLISA TruHits kit 20 ug/mL 
 
1.8E-02 >0.1 44

 

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 citrate using anti-6X His AlphaLISA Acceptor beads (with glutathione Donor beads) showed little effect on the assay signal at final citrate concentrations up to 0.77 mM. Citrate did show a half-maximal inhibition of signal when used at a final concentration of 0.94 mM. When 0.1 M citrate was included in the assay, 100% signal inhibition was observed.

 

citrate-and-bead-interference

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.

For research use only. Not for use in diagnostic procedures.