FlowCellect™ Bivariate Cell Cycle Kit for DNA Replication Analysis

FlowCellect™ Bivariate Cell Cycle Kit for DNA Replication Analysis

FlowCellect

25 Tests / Kit

Millipore’s FlowCellect™ Bivariate Cell Cycle Kit for DNA Replication Analysis is designed to investigate cells undergoing replication in S-phase of the cell cycle with high accuracy and confidence in flow cytometry applications. The kit includes a directly conjugated Anti-BrdU Alexa Fluor® 488 conjugate plus a DNA dye (Propidium Iodide) which allows the researcher to perform bivariate analysis to distinguish cells in two dimensions without the need of software modules for data interpretation. Flow cytometry allows the simultaneous measurement of incorporated BrdU as well as the DNA content on a single cell level. In this way, the cohort of labeled cells can be followed through the cell cycle.

The kit includes an optimized protocol with all of the necessary components to provide the researcher a true “plug and play” design for their research when studying DNA replication and its relation to cell proliferation and cancer. The kit includes a BrdU precursor to allow for BrdU measurement by the conjugated antibody and an RNase reagent to ensure that the PI intensity is directly proportional to DNA content only. This immunocytochemical method is very sensitive, and it offers an assay to screen in vivo inhibitors involved in the S-phase transition by flow cytometry. It is important to determine the biological characteristics of certain cancers, not only to evaluate their malignant potentials but also to decide on the most effective anticancer therapies.

Examination of the cell cycle has proven to be an accurate and effective method for assessing cell health by analyzing cell proliferation in great detail. Also recognized as the “Circle of Life”, the cell cycle can be divided into two distinct stages. The first stage is Interphase which consists of the G1, S, and G2 phases where cells are active, growing, and DNA replication takes place. The second is M-phase, also known as the “mitotic phase”, where cell division takes place. Researchers have employed various strategies to closely interrogate each of the critical steps at various phases in the cell cycle. For example, analysis of cells in S-phase will provide a direct measurement of newly synthesized DNA. Also, having the ability to discriminate cells between the G2 and M-phases can serve as a good indicator of cells undergoing mitosis. In all, a comprehensive understanding of cell cycle behavior can provide the researcher with useful information which will be important in interpreting the intrinsic nature of cell proliferation and assist in the development of anti-neoplastic agents.

Flow cytometric analysis of cell DNA content is widely used for the estimation of cell cycle phase distributions. However, the limitation in evaluating by only one marker (such as a DNA dye only) is that cells within each phase cannot accurately be determined without the use of software modules. Studying the cell cycle is now made easier by using a cell cycle phase-specific antibody plus a DNA dye in flow cytometry applications. Bivariate analysis using flow cytometry will not only reveal the distribution of cells in particular phases of the cycle, but will also allow the researcher to elucidate the molecular and functional mechanisms associated within the cell cycle.

Flow Cytometry

Human

• Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
• Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

Lot to lot in-process testing on all kit components

• 1. 20X Anti-BrdU - Alexa Fluor 488 Conjugated Monoclonal Antibody: (Part No. CS204818) One vial containing 150 µL antibody.
• 2. Propidium Iodide Solution: (Part No. CS204836) One vial containing 400 µL DNA dye.
• 3. BrdU (5-Bromo-2'-deoxyuridine) Reagent: (Part No. CS204820) One vial containing 100 µL.
• 4. RNase A Reagent: (Part No. CS204821) One vial containing 25 µL.
• 5. Fixation Buffer: (Part No. CS202122) One bottle containing 13 mL buffer.
• 6. 10X Wash Buffer: (Part No. CS202123) One bottle containing 13 mL buffer.
• 7. 5X Assay Buffer: (Part No. CS202124) One bottle containing 55 mL buffer.
• 8. 1X Permeabilization Buffer: (Part No. CS202127) One bottle containing 13 mL buffer.

Mouse

4-8°C for antibodies and buffers

Fluorescent

FlowCellect™ Bivariate Cell Cycle Kit for DNA Replication Analysis

Flow Cellect

Monoclonal Antibody

Research