Introduction And Product Description - PromoKine PK-CA724-488HTS Instruction Manual

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Instruction Manual

Introduction and product description:

The detection of cell proliferation is of utmost importance for assessing cell health,
determining genotoxicity or evaluating anticancer drugs. This is normally performed by
adding nucleoside analogs like [ H]thymidine or 5-bromo-2'-deoxyuridine (BrdU) to
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cells during replication, and their incorporation into DNA is detected or visualized by
autoradiography or with an anti-BrdU-antibody respectively. Both methods exhibit
several limitations. Working with [ H]thymidine is troublesome because of its
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radioactivity. Autoradiography is slow and thus not suitable for rapid high-throughput
studies. The major disadvantage of BrdU staining is that the double-stranded DNA
blocks the access of the anti-BrdU antibody to BrdU units. Therefore, samples have to
be subjected to harsh denaturing conditions resulting in degradation of the structure of
the specimen.
The PromoKine EdU-based Cell Proliferation Assays overcome these limitations,
providing a superior alternative to BrdU and [ H]thymidine assays for directly measuring
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DNA synthesis of adherent cells in 96 well plates. EdU (5-ethynyl-2'-deoxyuridine) is a
nucleoside analog to thymidine and is incorporated into DNA during active DNA
synthesis. In contrast to BrdU assays, the EdU-based Cell Proliferation Assays are not
antibody based and therefore do not require DNA denaturation for detection of the
incorporated nucleoside. Instead, the EdU-based Cell Proliferation Assays utilize click
chemistry for detection in a variety of dye fluorescent readouts. Furthermore, the
streamlined detection protocol reduces both the total number of steps and significantly
decreases the total amount of time. The simple click chemistry detection procedure is
complete within 30 minutes and is compatible with multiplexing for content and
context-rich results.
For research use only.
Information in this document is subject to change without notice. PromoCell GmbH
assumes no responsibility for any errors that may appear in this document.
PromoCell GmbH disclaims all warranties with respect to this document, expressed or
implied, including but not limited to those of merchantability or fitness for a particular
purpose. In no event shall PromoCell GmbH be liable, whether in contract, tort,
warranty, or under any statute or on any other basis for special, incidental, indirect,
punitive, multiple or consequential damages in connection with or arising from this
document, including but not limited to the use thereof.
Please read the material safety data sheets (MSDS) provided for each
product/component.
The EdU-Click technology is protected by patents WO2006/117161 and
WO 03/101972. Any commercial use will require a licence. Please contact us for details.
Also available: PromoKine EdU-based Cell Proliferation Assays available for Flow
Cytometry (PK-CA724-488FC, PK-CA724-555FC, PK-CA724-594FC & PK-CA724-
647FC) and Fluorescence Microscopy (PK-CA724-488FM, PK-CA724-555FM,
PK-CA724-594FM & PK-CA724-647FM) are also available.
See our website for more information.