Cellomics HCS Reagent Kit for mitotic activity indicator phospho-Histone H3.

Description

The Mitotic Index HCS Reagent Kit provides high-content screening (HCS)-quality fluorescence reagents and a validated protocol for measurement of mitotic index, or the fraction of cells undergoing cell division within a population. Cell division is a tightly regulated process involving complex sub-cellular changes, but two key mitotic events are easily identifiable: microtubule spindle formation and chromosome condensation. Chromatin condensation is initiated in late interphase by post-translational modification of the core histone proteins (Figure 1). Mitotic cells are identified selectively using an antibody specific for a phosphorylated epitope of a core histone protein, and a fluorescently labeled secondary antibody.

The Mitotic Index assay is used to characterize proliferating cells and to identify compounds that inhibit or induce mitotic progression. Compounds that inhibit mitotic progression result in an increase in the mitotic index of a population. Following compound treatment, cells growing in standard high density plates are fixed, permeabilized, and then labeled by indirect immunofluorescence. Prepared cells can be analyzed using standard fluorescence microscopy or using a Cellomics fully automated HCS Reader with Mitotic Index or Cell Cycle BioApplications, affording automated plate handling, focusing, image acquisition, analysis, quantification, and data storage.

Anti-mitotic drugs such as paclitaxel and vinblastine are used to treat various types of cancer. The Mitotic Index HCS Reagent Kit is a useful cell based assay tool for the identification of novel anti-mitotic compounds, and also as a means for diagnosing and characterizing cancerous cells. Skin fibroblasts from Alzheimer’s patients also show reduced proliferative capacity and mitotic index, broadening the applicability of the assay. Fully automated and validated screening for mitotic index in fields of whole cells is possible through the ArrayScan HCS Reader.

Features

• Cells prepared with the HCS Reagents can be analyzed directly via standard fluorescence microscopy; or HCS imaging systems
• Simple sample preparation/processing requires less than 2.5h after compound treatment
• Functional whole cell assay is more accurate and biologically relevant than flow cytometric analysis
• No cell lysis, purification, or filtration steps
• Kit antibody binds to epitope abundant in mitotic cells with high sensitivity and specificity

Related Poster Presentations

	V. C. Abraham, R. N. Ghosh, P. Sammak, P. Petrosko and J. R. Haskins (2004) Application of High Content Screening to the Analysis of Stem Cell Growth & Differentiation.
	Sarah Burroughs Tencza and Jeffrey R. Haskins (2004) High-Content Profiling of Cell Cycle Related Targets with RNAi.

Kit Contents

• Mitotic Index primary antibody
• Alexa Fluor® 488 conjugated secondary antibody
• Hoechst dye
• Positive control compound
• Wash buffer (10x)
• Permeabilization buffer (10x)
• Anti-Mitotic control compound (Vinblastine)
• Plate seals (10x)
• Block (100x)
  

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).

See all Cellomics HCS Reagent Kits