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Detecting intracellular activity is easier than ever, thanks to the HitKit series from Cellomics Inc. of Pittsburgh, Pa. Relying upon dyes and fluorophores visualized by fluorescence microscopy, the HitKit series of assay reagents permits visualization of various cellular events in intact cells. The kits are comprised of buffers, fluorescent probes, and sometimes positive or negative control compounds and are optimized to increase sensitivity, specificity, and signal-to-noise ratio. This optim

Apr 30, 2001
Kelly Karpa
Detecting intracellular activity is easier than ever, thanks to the HitKit series from Cellomics Inc. of Pittsburgh, Pa. Relying upon dyes and fluorophores visualized by fluorescence microscopy, the HitKit series of assay reagents permits visualization of various cellular events in intact cells.

The kits are comprised of buffers, fluorescent probes, and sometimes positive or negative control compounds and are optimized to increase sensitivity, specificity, and signal-to-noise ratio. This optimization, combined with each fluorophore's ability to absorb and emit light at distinct wavelengths, enables researchers to carry out multiplex assays. As such, the HitKit series of reagents is amenable to cell-based, high-content screening applications.

Four broad categories of cellular actions can be studied with the kits: indices of cell signaling, cell mitosis, cell motility and morphology, and cellular toxicity. A variety of kits are available for studying basic cell signaling properties involving transcription factor or kinase activation: NF-*B, STAT1, STAT2, STAT3, STAT5, c-Jun, ERK MAPK, p38 MAPK, and ATF-2. These transcription factors and kinases are primarily cytoplasmic until activated, when they translocate to the nucleus. Nuclear accumulation of these proteins may be observed using the kits. Analysis of compounds that induce or inhibit cell division may be performed with the Mitotic Index assay, in which mitotic cells are identified by detection of post-translationally modified core histone proteins. Three kits are available to measure cell motility and morphology, two useful indices for a variety of biological processes. The Cell Spreading kit quantifies cell attachment by measuring cellular areas, the Neurite Outgrowth kit identifies neurons within a heterogeneous mixture of cells, and the Cell Motility kit detects cellular movement by phagokinetic tracks generated during cellular migration. Cellular toxicity may also be analyzed by a variety of different parameters. The Cell Viability kit uses fluorescent probes to differentiate between live cells and dead cells, whereas the Multiparameter Apoptosis 1 kit measures nuclear morphology, mitochondrial mass, and changes in f-actin content simultaneously.

Andreas Vogt, research assistant professor in the department of Pharmacology at University of Pittsburgh, uses several products in the HitKit series: "They have really increased our productivity, allowing us to generate reliable, statistically meaningful data on hundreds of cells at the same time."

The broad range of reagent kits available make the HitKit series ideal for studying therapeutic applications such as inflammation, cancer, cell signaling pathways, cell stress, cardiovascular, and CNS processes. New kits for studying other aspects of cell signaling and cell toxicity are expected to debut soon.

-Kelly Karpa (kkarpa1@juno.com)

For More Information
Cellomics Inc.
(888) 826-3857
www.cellomics.com