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Chemi-Labeling Kits
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| Labeling
Kit |
| Format |
Catalog # |
Kit Insert |
MSDS |
Assay Worksheet
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| 96 Well Kit |
907-001 |
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FOR RESEARCH PURPOSES ONLY, NOT FOR DIAGNOSTIC USE. |
 
Direct Chemiluminescent Label
Label Proteins, Peptides or Nucleic Acids
Label, Purify and Detect in < 60 minutes
Detect less than 1 attomole (10-18) of IgG
NO Enzymes or Radioactivity
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The Assay Designs' Sub-Attomole Labeling Kit is designed to allow the
rapid attachment of a chemiluminescent Acridinium Ester to lysine groups
on most antibodies, proteins, nucleic acids and some peptides. The kit
contains enough material to perform up to 5 labeling experiments
equivalent to 1 mg of IgG each. The following materials are included:
Acridinium Ester with a N-hydroxy- succinimide Ester group, dry solvent
for dissolving the Acridinium Ester, buffer for diluting the antibody,
protein or nucleic acid, a solution of lysine for stopping the labeling
reaction, a gel filtration column for separation of the labeled product
from excess Acridinium Ester, column running buffer, and a set of
Trigger solutions for detection of the labeled antibody, protein or
nucleic acid.
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Acridinium Ester labeled samples can be used for a variety of different
applications. They can be added to samples containing the unlabeled
materials as an alternative to the use of 125I labeled tracer
molecules. Acridinium Esters can be used to label antigens for
competitive immunoassays, or signal antibodies for immunometric assays.
In addition, chemiluminescent labeled nucleic acids may be used as
reporter molecules in DNA probe based assay systems.
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The Acridinium Ester supplied in this kit has a N-hydroxysuccinimidyl (NHS)
ester labeling group attached to a 2 carbon spacer arm. The NHS ester
labeling group will covalently attach to any primary amino group on the
protein, peptide or nucleic acid to be labeled. At pH's above neutral,
the NHS ester labeling group is subject to nucleophilic attack by amine
groups on the protein, peptide or nucleic acid. As a result, the NHS
group is displaced from the Acridinium Ester to form a stable amide bond
between the Acridinium Ester and the protein, peptide or nucleic acid.
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Acridinium C2 N-Hydroxysuccinimide Ester
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The following data is 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 were
compared to two (2) standard deviations from the zero. In this case the
lowest amount of mouse IgG that could be detected was
0.6 x 10-18 Moles.
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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 Service 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 yield
greater sensitivity, speed of analysis, dynamic range and/or decreased
sample volume. Assay Designs has published papers on the use of
Acridinium Esters to detect prostaglandins with speed and sensitivity in
excess of traditional immunoassay methods. |
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