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Acridinium C(2)
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Luminescent Products |
| Product |
Catalog # |
Kit Insert |
MSDS |
| Acridinium C2 NHS Ester |
906-000 |
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| Trigger Solutions |
906-001 |
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FOR RESEARCH PURPOSES ONLY, NOT FOR DIAGNOSTIC USE. |
 
Simple NHS Chemistry
Direct, High Specific Activity Chemiluminescent Label
Labels Any Primary Amine Group
Can Be Read in Most Plate or Tube Luminometers
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Chemiluminescence is the emission of light from a chemical reaction that
occurs at or near ambient temperatures. A vast number of reactions give
rise to the emission of light in solution, but only a few have
sufficiently high efficiencies of chemiluminescence to have been used
for analytical purposes. For further information on the vast array of
chemical reactions that produce light please refer to excellent reviews
by McCapra and Beheshti in "Bioluminescence and Chemiluminescence:
Instruments and Applications", 1985, Ed. K. Van Dyke, CRC Press, Boca
Raton, FL. pgs. 9-42; and by Schuster and Smith, "Adv. Phys. Org. Chem.",
1982, 18, 187.
Acridinium esters have moderate chemiluminescence efficiencies and
the chemistry of light production is well understood. They have the
added benefit that they require simple triggering conditions that
contribute little to the background signal. This benefit has been
exploited to great effect in high sensitivity immunoassay applications.
These compounds can also be easily modified to attach to proteins and
other molecules of interest, without significantly affecting their
chemiluminescence properties.
Attachment to proteins, antibodies, peptides or nucleic acids
containing a primary amino group is carried out by dissolving the
Acridinium ester in a dry aprotic solvent such as dimethyl formamide and
adding the solution to the material to be labeled in a suitable buffer.
Suitable buffers are ones without amine groups, such as borate or
bicarbonate buffers at pH's between 7 and 10. We suggest labeling at pH
8.5 for most reactions. Excess Acridinium ester label is removed by
dialysis or by gel permeation chromatography through a resin such as
Sephadex G-10.
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The Acridinium Ester labeled samples can be used for a number of
different applications. They can be added as tracers to samples
containing unlabeled materials as an alternative to the use of 125I
labeled molecules. They can be used as labeled antigens for competitive
immunoassays, and as labeled signal antibodies for immunometric assays.
Chemiluminescent labeled nucleic acids may be used as reporter molecules
in DNA probe based systems. |
The following data is a typical column purification for the Acridinium
Ester labeling of mouse IgG. The graph shows the Relative Light Units (RLU)
for each fraction.
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The fraction containing the Acridinium Ester-labeled mouse IgG was used
for the sensitivity determination. The labeled protein was serially
diluted from 1:100 to 1:106 in Column Buffer and duplicate
measurements were made in a Berthold tube luminometer. Acridinium Ester
chemiluminescence takes 2 seconds using the Trigger Solutions provided.
Sensitivity was calculated by determining the RLU's emitted from the
buffer blank and comparing to the RLU's emitted by the lowest
concentration of labeled mouse IgG. Here, the difference in RLU's was
compared at two (2) standard deviations from the zero. Under these
conditions, the lowest amount of mouse IgG that could be detected was
0.6 x 10-18 Moles. |
The uses of Acridinium Ester labeled antibodies,
antigens and DNA have been described in a number of publications.
These references are available upon request from the Assay
Designs' Technical Services Team.
Streptavidin, progesterone, thyroxine and free thyroxine,
alpha-fetoprotein, thyroid stimulating hormone, human chorionic
gonadotropin, human complement component C9, staphylococcal
peptidoglycan, carcinoembryonic antigen, Chlamydia
trachomatis and Neisseria gonorrhoeae RNA have all
been detected using this molecule or analogs. Invariably the
assays developed using this material show greater sensitivity,
speed of analysis, dynamic range and/or decreased sample volume.
| Material |
Catalog Number |
Quantity |
| Acridinium C2 NHS Ester |
906-000
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1 mg |
| Acridinium Trigger Solutions |
906-001
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1 set, 250 mL each |
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+32 (0) 16
58 90 45 
+32 (0) 16
50 90 45