FlowCellect Mouse ESC Nuclear Marker Characterization Kit

FlowCellect Mouse ESC Nuclear Marker Characterization Kit

FlowCellect

Millipore’s The FlowCellect™ Mouse Embryonic Stem Cell Characterization Kit is designed as an easier and quicker way to track surface marker expression of mOCT4, SSEA4 and SSEA1. The kit will also help to determine the percentage of undifferentiated mouse embryonic stem cells in culture by determining the percentage of cells that express both OCT4 and SSEA1 and not SSEA4. This quick test will allow researchers to determine the quality of their cells in culture as well as see changes in marker expression during a differentiation protocol.
Sufficient reagents are provided for 25 3-color samples and 5 isotype control tests. The kit includes all optimized fluorescently labeled antibodies and buffers necessary for analysis. A cell strainer is also included to ensure samples are devoid of large cell clumps that can alter cell analysis. Detailed assay instructions are also included to assist in analysis and to ensure the correct cell concentration is obtained for flow cytometric analysis.

Mouse embryonic stem cells (mESCs) are unique cells in that they can be maintained in vitro as pluripotent cells and can be induced to differentiate into many cell types by manipulating culture conditions. mESCs have been used as a model to help define differentiation and signaling pathways in human embryonic stem cells (hESCs), since hESCs are more arduous to work on. Therefore, much attention has been focused on understanding the mechanisms behind mESC differentiation.

Currently, the most utilized method to test the differentiation state of mESCs is to analyze the expression of the proteins OCT4 and the stage specific embryonic antigen, SSEA1 (1,2). The transcription factor, OCT4, is highly expressed in undifferentiated mESCs and is essential for the formation and maintenance of pluripotentcy (3,4). SSEA1 is also expressed on undiiferentiated mESCs but another stage specific embryonic antigen, SSEA4, is not (2,5,6). However, upon retinoic treatment, SSEA1 expression wanes and in some instances, accompanied by expression of SSEA4 (6,7).

The FlowCellect™ Mouse Embryonic Stem Cell Characterization Kit uses an antibody-based detection method, utilizing fluorescently conjugated antibodies to detect OCT4, SSEA4 and SSEA1. The FlowCellect™ Mouse Embryonic Stem Cell Characterization kit is designed specifically for flow cytometers with a single blue (488 nm) laser light source. The FlowCellect™ Mouse Embryonic Stem Cell Characterization Kit is not intended for use in microscopy applications including ICC, IHC and HTS, western blot or any other fluorometric analysis platforms.

The kit is optimized for use on the Guava EasyCyte™ Plus cytometer that requires cell concentrations to be less than 500 cells per microliter. The kit does not include culture media or other materials that should be readily available in a common laboratory setting that routinely performs experiments of the type described here.

• Flow Cytometry
• in vitro Toxicology Assays

Mouse

• 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

Stem Cell Kits

Mouse Embryonic Stem Cells

Stem Cell Research

Stem Cell

Characterization

• 1. 10X Wash buffer: (Part No. CS202123) 1 bottle containing 13mL
• 2. Fixation buffer: (Part No. CS202122) 1 bottle containing 13mL
• 3. 5X Assay buffer: (Part No. CS202124) 1 bottle containing 55mL
• 4. 1X Permeabilization Buffer: (Part No. CS202125) 1 bottle containing 13mL
• 5. 20X rat IgG2a-Alexa 488 isotype control: (Part No. CS202404) 1 vial containing 75uL
• 6. 20X IgG3-PE isotype control: (Part No. CS202405) 1 vial containing 75uL
• 7. 20X IgM-PE/CY5 isotype control: (Part No. CS202397) 1 vial containing 75uL
• 8. 20X mOCT4-Alexa 488: (Part No. CS202394) 1 vial containing 150uL
• 9. 20X SSEA4-PE: (Part No. CS202399) 1 vial containing 150uL
• 10. 20X SSEA1-PE/CY5: (Part No. CS202400) 1 vial containing 150uL
• 11. Steri-flip cell strainer (60mm): (Catalog # SCNY00060)

4-8°C; 6 Months

Mouse

Fluorescent

• FUT4
• SSEA-4

Flow Cellect

ExpressPro

Monoclonal Antibody

Research