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AlphaLISA AlphaScreen

Urea

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
Urea
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 urea might interfere in assays involving particular bead products, and at what urea 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 

AL126 

Protein L AlphaLISA Acceptor Beads 

>0.1 

>0.1 

AL127 

Anti-FITC AlphaLISA Acceptor Beads 

3.8E-02 

>0.1 

21 

AL128 (biotin) 

Anti-6xHis AlphaLISA Acceptor Beads 

9.3E-02 

>0.1 

13 

AL128 (GSH) 

Anti-6XHis AlphaLISA Acceptor Beads 

2.0E-02 

>0.1 

19 

AL129 

Anti-V5 AlphaLISA Acceptor Beads 

>0.1 

>0.1 

AL130 

Anti-mouse IgM AlphaLISA Acceptor Beads 

>0.1 

>0.1 

AL131 

Anti-chicken IgY AlphaLISA Acceptor Beads 

>0.1 

>0.1 

11 

AL132 

Anti-sheep IgG AlphaLISA Acceptor Beads 

>0.1 

>0.1 

AL133 

Anti-GFP AlphaLISA Acceptor Beads 

>0.1 

>0.1 

AL134 

Anti-MBP AlphaLISA Acceptor Beads 

>0.1 

>0.1 

AL136 

Strep-Tactin AlphaLISA Acceptor Beads 

>0.1 

>0.1 

AS102 

Protein A Alpha Donor Beads 

>0.1 

>0.1 

AS103 

Anti-FLAG Alpha Donor Beads 

>0.1 

>0.1 

AS104 

Anti-mouse IgG Alpha Donor Beads 

>0.1 

>0.1 

AS105 

Anti-rabbit IgG Alpha Donor Beads 

>0.1 

>0.1 

AS106 

Strep-Tactin Alpha Donor Beads 

>0.1 

>0.1 

AL900 

AlphaLISA TruHits kit 2 ug/mL 

>0.1 

>0.1 

AL900 

AlphaLISA TruHits kit 20 ug/mL 

>0.1 

>0.1 

 

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 urea using anti-6X His AlphaLISA Acceptor beads (with glutathione Donor beads) showed little effect on the assay signal at final urea concentrations up to 20 mM. When 0.1 M urea was included in the assay, 19% signal inhibition was observed.

urea-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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