Magnesium 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 Mg2+ might interfere in assays involving particular bead products, and at what Mg2+ 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 | >0.1 | >0.1 | 0 |
| AL126 | Protein L AlphaLISA Acceptor Beads | >0.1 | >0.1 | 0 |
| AL127 | Anti-FITC AlphaLISA Acceptor Beads | 1.6E-02 | 7.3E-02 | 60 |
| AL128 (biotin) | Anti-6xHis AlphaLISA Acceptor Beads | 1.2E-02 | 5.3E-02 | 72 |
| AL128 (GSH) | Anti-6XHis AlphaLISA Acceptor Beads | 1.8E-03 | 2.6E-02 | 86 |
| AL129 | Anti-V5 AlphaLISA Acceptor Beads | >0.1 | >0.1 | 0 |
| AL130 | Anti-mouse IgM AlphaLISA Acceptor Beads | 7.6E-03 | 3.1E-02 | 63 |
| AL131 | Anti-chicken IgY AlphaLISA Acceptor Beads | 3.6E-02 | >0.1 | 20 |
| AL132 | Anti-sheep IgG AlphaLISA Acceptor Beads | 1.0E-02 | >0.1 | 26 |
| AL133 | Anti-GFP AlphaLISA Acceptor Beads | >0.1 | >0.1 | 0 |
| AL134 | Anti-MBP AlphaLISA Acceptor Beads | 3.2E-02 | >0.1 | 32 |
| AL136 | Strep-Tactin AlphaLISA Acceptor Beads | 3.4E-02 | >0.1 | 44 |
| AS102 | Protein A Alpha Donor Beads | >0.1 | >0.1 | 0 |
| AS103 | Anti-FLAG Alpha Donor Beads | >0.1 | >0.1 | 0 |
| AS104 | Anti-mouse IgG Alpha Donor Beads | 3.2E-02 | 5.1E-02 | 52 |
| AS105 | Anti-rabbit IgG Alpha Donor Beads | >0.1 | >0.1 | 0 |
| AS106 | Strep-Tactin Alpha Donor Beads | >0.1 | >0.1 | 0 |
| AL900 | AlphaLISA TruHits kit 2 ug/mL | >0.1 | >0.1 | 17 |
| 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 MgCl2 using anti-6X His AlphaLISA Acceptor beads (with glutathione Donor beads) showed little effect on the assay signal at final MgCl2 concentrations up to 1.8 mM. MgCl2 did show a half-maximal inhibition of signal when used at a final concentration of 26 mM. When 0.1 M MgCl2 was included in the assay, 86% signal inhibition was observed.
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.
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- General Page on Buffer Considerations
- 2-Mercaptoethanol Interference
- Adenosine (ATP) Interference
- CHAPS Interference
- Citrate Interference
- Deoxycholate Interference
- DMSO Interference
- DTT Interference
- EDTA Interference
- EGTA Interference
- Gelatin Interference
- Glycerol Interference
- Glycine Interference
- IBMX Interference
- Imidazole Interference
- Magnesium Interference
- Nitroprusside Interference
- Roche Complete Protease Inhibitor Interference
- S-Adenosylmethionine (SAM) Interference
- SDS Interference
- Sodium Fluoride (NaF) Interference
- Tris Interference
- Triton X-100 Interference
- Tween 20 Interference
- Urea Interference
- Vanadate Interference