Cellomics HCS Reagent Kits for multiplex analysis of cell proliferation parameters.

Description

The BrdU and Ki67 Cell Proliferation HCS Reagent Kits are for simultaneous quantification of DNA replication and Ki67 proliferation marker in the same cell. These kits allow direct measurements of BrdU incorporation and Ki67 expression change using a fixed end-point assay based on immunofluorescence detection in cells grown on standard high-density microplates. The primary antibodies are specific for their targets and have minimal cross-reactivity with other targets.

The green BrdU and Orange Ki67 multiplex kits contain primary antibodies towards BrdU and Ki67 (mouse monoclonal and rabbit polyclonal, respectively) and secondary antibodies conjugated with green and orange fluorophores (DyLight™ 488 Goat anti-Mouse antibody and DyLight™ 549 Goat anti-Rabbit antibody, respectively). The orange BrdU singleplex kit contains a mouse monoclonal primary antibody towards BrdU and DyLight™ 549 Goat anti-Mouse antibodies. The orange Ki67 probe kit contains a rabbit polyclonal primary antibody towards Ki67 and a DyLight™ 549 Conjugated Goat anti-Rabbit secondary antibody.

The BrdU and Ki67 Cell Proliferation HCS Reagent Kits have been optimized for use with the Cellomics ArrayScan HCS Reader using the Target Activation BioApplication Software Module. These kits can be used with other Cellomics BioApplications (see the Compatible BioApplication Software Modules Section). Thus, automated plate handling, focusing, cell image acquisition/processing, and data analysis/management are combined in one high content screening (HCS) system to assay for test compounds regulating BrdU incorporation and the level of Ki67 nuclear antigen. This assay is sensitive, rapid and easy to use.  The evaluation of cell proliferation as well as other information such as cell number, morphology and analysis of other cellular antigens can be obtained from a single culture. In addition to HCS instruments, cells labeled by the reagents in this kit can be viewed and analyzed by other fluorescence microscopy-based instruments.

Cell proliferation and the characterization of agents that either promote or inhibit cell proliferation are particularly critical areas of cell biology and drug-discovery research. Although cell proliferation can be measured by the incorporation of [³H]-thymidine into cellular DNA, this assay is slow, labor intensive and its use is limited by potentially hazardous radioactive materi­als. As an alternative to [³H]-thymidine, 5-bromo-2´-deoxyuridine (BrdU), a thymidine analog, can be used to detect the DNA replication in actively proliferating cells. Cells which have incorporated BrdU into DNA can be quickly detected by using monoclonal antibodies directed against BrdU and fluorophore-conjugated secondary antibody. BrdU staining facilitates the identification of cells that have progressed through the S-phase of the cell cycle during the BrdU-labeling period. Several protein markers are known to be associated with cell proliferation, including the antigen Ki67 which is expressed during G1, S, G2, and M phases of the cycle but absent in G0 phase, such as in quiescent and terminally differentiated cells. Ki67 antigen-positive cells give a more specific and accurate indication of proliferating cells compared to that of PCNA-positive cells since PCNA is detectable in almost all quiescent cells. Ki67 has also been found to correlate with poor survival in ovarian cancer in both univariate and multivariate analyses. The quantification of Ki-67 provides independent prognostic information in breast cancer in addition to the tumor size and S-phase fraction.

Related Poster Presentations

	Suk J. Hong, Richik N. Ghosh (2007) A Novel High Content Screening Assay for Monitoring Cell Proliferation Activity by Multiplexed Measurements of DNA Content, BrdU and Ki67.
	Suk J. Hong, Kay Opperman, Richik N. Ghosh (2007) New Reagent Kits for the High Content Analysis of Cell Proliferation Activity using BrdU Incorporation and Ki67 Expression.
	Sarah Burroughs Tencza and Jeffrey R. Haskins (2004) High-Content Profiling of Cell Cycle Related Targets with RNAi.

Ordering Information

Screening size kits and components (bulk and custom) available with special pricing. Please call.

To order Cellomics HCS Reagent Kits, please call 800-874-3723 in the U.S.
or contact your local distributor of Thermo Scientific Products.

Choose "Order Online" in table to see pricing and order online (U.S. only).

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