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AlphaLISA AlphaScreen

Anti-rabbit IgG Antibody-coated Donor Beads

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

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 what buffer components might interfere with AlphaLISA assays using Anti-Rabbit IgG Alpha Donor 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% loss Tested up to % of inhibition at max conc
Glycerol % 0.2 3.9 30% 91
DMSO % 0.5 1.8 10% 87
Gelatin % >1 >1 1% 0
CHAPS % 0.04 0.1 3% 84
Deoxycholate % 0.005 0.02 3% 94
SDS % 0.07 0.1 3% 96
Triton X-100 % >3 >3 3% 0
Tween 20 % >3 >3 3% 0
TRIS (pH 8.0) M 5.2E-02 4.5E-01 0.5M 53
2-Mercaptoethanol M 5.6E-04 1.1E-02 0.1M 82
DTT M 4.9E-08 6.1E-03 0.1M 75
EDTA M 4.3E-02 >0.1 0.1M 38
EGTA M 6.6E-03 >0.1 0.1M 34
IBMX M 1.0E-06 >0.0025 0.0025M 37
Imidazole M 1.3E-03 7.9E-03 0.1M 99
Mg2+ (MgCl2) M >0.1 >0.1 0.1M 0
Nitroprusside M 2.6E-03 2.9E-02 0.1M 65
Urea M >0.1 >0.1 0.1M 0
Vanadate M 9.8E-03 1.1E-02 0.1M 43
Citrate M 1.7E-03 3.9E-03 0.1M 100
Adenosine (ATP) M >0.01 >0.01 0.01M 0
Glycine M 1.9E-06 >0.01 0.01M 15
NaF M >0.01 >0.01 0.01M 0
S-adenosylmethionine M >0.001 >0.001 0.001M 0
Complete Protease Inhibitor X not determined not determined 1X 29

 

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-Rabbit IgG Donor beads (with Streptavidin AlphaLISA Acceptor beads) showed little effect on the assay signal at final Triton X-100 concentrations up to 3%. Using 3.9% glycerol resulted in a 50% loss of signal.

interferencetesting_ask.jpeg

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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