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AlphaLISA AlphaScreen

IBMX

Section
Bead Selection and Bead Interference
Alpha Antibody Detection and Characterization
Alpha Bead Conjugation
Alpha Citations
Alpha EMSA Conversion
Alpha Immunogenicity
Alpha Instrument Options and Settings
Alpha Products and Catalog Numbers
Alpha Protein-protein and Protein-nucleic Acid Interactions
Alpha Troubleshooting Tables
AlphaLISA 5 μL vs. 20 μL Sample Volume
AlphaLISA Epigenetic Toolbox Reagents
AlphaLISA Immunoassay Kits
AlphaLISA and AlphaScreen No-wash Assays
Bead Selection and Bead Interference
Buffer Selection for Alpha Assays
Create your own Alpha Assay
Data Analysis for AlphaLISA Immunoassays
Determining Kd With an Alpha Assay
Other Alpha Applications
Preparation of Analyte-depleted Serum for Alpha Assays
The Hook Effect
Working with Serum and Other Biological Matrices in Alpha Assays
Working with cell extracts and supernatants in Alpha assays
Sub Section
IBMX
2-Mercaptoethanol
Adenosine (ATP)
Anti-6X His antibody-coated AlphaLISA Acceptor Beads
Anti-FITC antibody-coated AlphaLISA Acceptor Beads
Anti-FLAG Antibody-coated Alpha Donor Beads
Anti-GFP Antibody-coated AlphaLISA Acceptor Beads
Anti-V5 Antibody-coated AlphaLISA Acceptor Beads
Anti-chicken IgY Antibody-coated AlphaLISA Acceptor Beads
Anti-maltose Binding Protein (MBP) Antibody-coated AlphaLISA Acceptor Beads
Anti-mouse IgG Antibody-coated Donor Beads
Anti-mouse IgM Antibody-coated AlphaLISA Acceptor Beads
Anti-rabbit IgG Antibody-coated Donor Beads
Anti-sheep IgG Antibody-coated AlphaLISA Acceptor Beads
CHAPS
Citrate
DMSO
DTT
Deoxycholate
EDTA
EGTA
Gelatin
Glutathione AlphaLISA Acceptor Beads
Glycerol
Glycine
IBMX
Imidazole
Magnesium and bead interference
Nickel Chelate AlphaLISA Acceptor Beads
Nitroprusside
Protein A-coated Donor Beads
Protein L-coated AlphaLISA Acceptor beads
Roche Complete Protease Inhibitor
S-adenosylmethionine (SAM)
SDS
Sodium Fluoride (NaF)
Strep-Tactin-coated Beads
Streptavidin-coated AlphaLISA Acceptor beads
TRIS pH 8.0
Triton X-100
Tween 20
Urea
Vanadate

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 IBMX might interfere in assays involving particular bead products, and at what IBMX 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 0.0025M 
Catalog Number  Product  No effect (10%)  50% loss  % of inhibition at max conc 
AL125  Streptavidin AlphaLISA Acceptor Beads  1.0E-06  >0.0025  37 
AL126  Protein L AlphaLISA Acceptor Beads  0.0E+00  >0.0025  35 
AL127  Anti-FITC AlphaLISA Acceptor Beads  1.1E-03  >0.0025  42 
AL128 (biotin)  Anti-6xHis AlphaLISA Acceptor Beads  >0.0025  >0.0025 
AL128 (GSH)  Anti-6XHis AlphaLISA Acceptor Beads  >0.0025  >0.0025 
AL129  Anti-V5 AlphaLISA Acceptor Beads  5.3E-06  1.5E-04  59 
AL130  Anti-mouse IgM AlphaLISA Acceptor Beads  1.2E-05  4.2E-05  53 
AL131  Anti-chicken IgY AlphaLISA Acceptor Beads  1.4E-03  >0.0025  39 
AL132  Anti-sheep IgG AlphaLISA Acceptor Beads  1.0E-03  >0.0025  46 
AL133  Anti-GFP AlphaLISA Acceptor Beads  2.5E-04  3.3E-04  59 
AL134  Anti-MBP AlphaLISA Acceptor Beads  2.4E-04  >0.0025  25 
AL136  Strep-Tactin AlphaLISA Acceptor Beads  >0.0025  >0.0025 
AS102  Protein A Alpha Donor Beads  6.4E-04  >0.0025  48 
AS103  Anti-FLAG Alpha Donor Beads  >0.0025  >0.0025 
AS104  Anti-mouse IgG Alpha Donor Beads  1.0E-03  >0.0025  40 
AS105  Anti-rabbit IgG Alpha Donor Beads  1.0E-06  >0.0025  37 
AS106  Strep-Tactin Alpha Donor Beads  >0.0025  >0.0025 
AL900  AlphaLISA TruHits kit 2 ug/mL  >0.0025  >0.0025 
AL900  AlphaLISA TruHits kit 20 ug/mL  2.2E-03  >0.0025  24 

 

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 IBMX using anti-6X His AlphaLISA Acceptor beads (with glutathione Donor beads) showed little effect on the assay signal at final IBMX concentrations up to 2.5 mM. 

ibmx-and-bead-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.

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