Cell Cycle Kit I

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 Cell Cycle Kit I
 Product# Price  Product Name  Pack Size  
         
 8404601      -    Cell Cycle I  1 x 96    SELECT   
 8404602      -    Cell Cycle I  5 x 96    SELECT   
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  Details All Product Numbers
 

Figure 3. Activation or inhibition of BrdU and phospho-histone H3 after drug treatment for 24 hours (top). Cell cycle drugs were added to the cells at the indicated concentrations (bottom). BrdU was measured using the output parameter of the nuclear fluorescence total intensity (MEAN_TotalIntenCh2) and phospho-histone H3 was measured using the nuclear fluorescence average intensity (MEAN_AvgIntenCh3) with the Cellomics Compartmental Analysis Bioapplication. Values are normalized with the maximum control value and presented as percent maximum in top image and percent control in bottom image.

 
Figure 2. BrdU and phospho-Histone H3 staining with cell cycle inhibitory drugs. A549 cells were treated with hydroxyurea (10 mM), nocodazole (0.5 ?g/ml), vinblastine (0.3 ?M), cycloheximide (10 ?g/ml), mimosine (0.5 mM) and sodium butyrate (5 mM) for 24 hours and stained with BrdU and phospho-Histone H3 antibodies as described in the protocol. Images of nuclei, BrdU and phoshpo-Histone H3 in drug-treated A549 cells.

These kits have been optimized with the Cellomics ArrayScan HCS Reader using the Target Activation BioApplication Software Module. Thus, automated plate-handling, focusing, cell image acquisition and processing, and data analysis are combined in one high-content screening (HCS) system to assay for test compounds. In addition to HCS instruments, cells labeled by the kit reagents can be viewed and analyzed by fluorescence and confocal microscopes.

The Cell Cycle Kit I BrdU and Phospho-Histone H3 Kit is for simultaneous quantification of nuclear DNA content to distinguish 2N (G1 phase) and 4N (G2/M phase), DNA replication in S phase cells, and mitosis marker in M phase cells. These kits allow direct measurements of BrdU incorporation and mitosis-specific histone H3 phosphorylation using a fixed end-point assay based on immunofluorescence detection in cells grown on standard high-density microplates. The DNA binding dye DAPI is used to determine the cell cycle phases by DNA content. The primary antibodies are specific for their targets and have minimal cross-reactivity with other targets.

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The Cell Cycle Kit I detects BrdU and phospho-Histone H3 in nuclear DNA without harsh treatments, enabling multiplexed detection of BrdU with antibodies toward other cellular targets. This assay is sensitive, rapid and easy to use. The evaluation of DNA replication, mitosis and other information such as cell number, the state of cell cycle-associated proteins, morphology and analysis of other cellular antigens can be obtained from a single culture.