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Imidazole 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 imidazole might interfere in assays involving particular bead products, and at what imidazole 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 

1.3E-03 

7.9E-03 

99 

AL126 

Protein L AlphaLISA Acceptor Beads 

7.8E-04 

6.2E-03 

99 

AL127 

Anti-FITC AlphaLISA Acceptor Beads 

1.3E-03 

5.9E-03 

100 

AL128 (biotin) 

Anti-6xHis AlphaLISA Acceptor Beads 

1.1E-03 

3.4E-03 

100 

AL128 (GSH) 

Anti-6XHis AlphaLISA Acceptor Beads 

6.6E-04 

2.4E-03 

100 

AL129 

Anti-V5 AlphaLISA Acceptor Beads 

4.6E-04 

3.3E-03 

99 

AL130 

Anti-mouse IgM AlphaLISA Acceptor Beads 

1.3E-03 

6.3E-03 

100 

AL131 

Anti-chicken IgY AlphaLISA Acceptor Beads 

9.2E-04 

5.3E-03 

99 

AL132 

Anti-sheep IgG AlphaLISA Acceptor Beads 

9.3E-04 

5.3E-03 

99 

AL133 

Anti-GFP AlphaLISA Acceptor Beads 

1.0E-03 

4.7E-03 

100 

AL134 

Anti-MBP AlphaLISA Acceptor Beads 

2.1E-03 

6.3E-03 

99 

AL136 

Strep-Tactin AlphaLISA Acceptor Beads 

1.3E-03 

6.8E-03 

99 

AS102 

Protein A Alpha Donor Beads 

5.9E-04 

4.8E-03 

99 

AS103 

Anti-FLAG Alpha Donor Beads 

1.1E-03 

6.2E-03 

99 

AS104 

Anti-mouse IgG Alpha Donor Beads 

6.9E-04 

5.0E-03 

98 

AS105 

Anti-rabbit IgG Alpha Donor Beads 

1.3E-03 

7.9E-03 

99 

AS106 

Strep-Tactin Alpha Donor Beads 

8.7E-04 

5.1E-03 

100 

AL900 

AlphaLISA TruHits kit 2 ug/mL 

9.1E-04 

5.7E-03 

99 

AL900 

AlphaLISA TruHits kit 20 ug/mL 

1.0E-03 

5.9E-03 

99 

 

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 imidazole using anti-6X His AlphaLISA Acceptor beads (with glutathione Donor beads) showed little effect on the assay signal at final imidazole concentrations up to 0.66 mM. Imidazole did show a half-maximal inhibition of signal when used at a final concentration of 2.4 mM. When 0.1 M imidazole was included in the assay, 100% signal inhibition was observed.

 

imidazole-interference-fig1.png

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.