EasySep™ Mouse Mammary Stem Cell Enrichment Kit

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Immunomagnetic Negative Selection of Mouse Mammary Stem Cells (MaSC)

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  • Test Tube Box EasySep®
  • Box top label for Mouse Mammary Stem Cell Enrichment Kit For labeling 10e9 cells
Test Tube Box EasySep®
The EasySep™ Mouse Mammary Stem Cell Enrichment Kit is designed to isolate mammary stem cells from freshly dissociated mouse mammary tissues. Unwanted cells are targeted for removal with biotinylated antibodies that are directed against non-epithelial cells (CD45, CD31, TER119, BP-1). Labeled cells are then recognized by Tetrameric Antibody Complexes that are directed against biotin and dextran. These cells are bound to magnetic particles and separated using an EasySep™ magnet without the use of columns. The untouched epithelial cell-enriched fraction is poured off into a new tube and then labeled with CD24 and CD49f antibodies so that the CD24+CD49f++ sub-population can be isolated by fluorescence-activated cell sorting.
Product Name Description Catalog # Size Price Quantity
EasySep™ Mouse Mammary Stem Cell Enrichment Kit Immunomagnetic negative selection of mouse mammary stem cells (MaSC) 19757 For labeling up to 1 x 10e9 cells 998.00 USD      
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Recommended for:
The isolation of mouse mammary stem cells from freshly dissociated mammary tissue by negative selection
Components:
• EasySep™ Mouse Epithelial Cell Enrichment Cocktail, 0.5 mL
• EasySep™ Biotin Selection Cocktail, 1.0 mL
• EasySep™ Magnetic Particles, 1.0 mL
• CD24 Antibody, Clone M1/69, PE-Conjugated, 0.5 mL
• CD49f Antibody, Clone GoH3, FITC-Conjugated, 1.0 mL
Accessory Products:
• Collagenase/Hyaluronidase Solution (Catalog #07912)
• Gentle Collagenase/Hyaluronidase Solution (Catalog #07919)
Intended Use Statement: For Research Use Only. Not for Therapeutic or Diagnostic Use.
Contains:
N/A
Equipment Required:
• EasySep™ Magnet (Catalog #18000), or
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
Product Type: Cell isolation products
Area of Interest: Mammary cell research
Cell Isolation Product Type: Reagents
Cell Source: Other
Cell Type: Mammary epithelial cells
Popular Product Line: EasySep
Selection Method: Negative
Species: Mouse

Procedures and instruction manuals:

Educational resources:

MSDS:

FAQS:


  • HOW EASYSEP™ WORKS

    • Q. CAN EASYSEP™ BE USED FOR EITHER POSITIVE OR NEGATIVE SELECTION?
      A. Yes. The EasySep™ kits use either a negative selection approach by targeting and removing unwanted cells or a positive selection approach targeting desired cells. Depletion kits are also available for the removal of cells with a specific undesired marker (e.g. GlyA).
    • Q. HOW DOES THE SEPARATION WORK?
      A. Magnetic particles are crosslinked to cells using Tetrameric Antibody Complexes (TAC). When placed in the EasySep™ Magnet, labeled cells migrate to the wall of the tube. The unlabeled cells are then poured off into a separate fraction.
    • Q. WHICH COLUMNS DO I USE?
      A. The EasySep™ procedure is column-free. That's right - no columns!
    • Q. HOW CAN I ANALYZE THE PURITY OF MY ENRICHED SAMPLE?
      A. The Product Information Sheet provided with each EasySep™ kit contains detailed staining information.
    • Q. CAN EASYSEP™ SEPARATIONS BE AUTOMATED?
      A. Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.
    • Q. CAN EASYSEP™ BE USED TO ISOLATE RARE CELLS?
      A. Yes. We recommend a cell concentration of 2x108 cells/mL and a minimum working volume of 100 µL. Samples containing 2x107 cells or fewer should be suspended in 100 µL of buffer.

    EASYSEP™ MAGNETIC PARTICLES

    • Q. ARE THE EASYSEP™ MAGNETIC PARTICLES FACS-COMPATIBLE?
      A. Yes, the EasySep™ particles are flow cytometry-compatible, as they are very uniform in size and about 5000X smaller than other commercially available magnetic beads used with column-free systems.
    • Q. CAN THE EASYSEP™ MAGNETIC PARTICLES BE REMOVED AFTER ENRICHMENT?
      A. No, but due to the small size of these particles, they will not interfere with downstream applications.
    • Q. HOW DOES THE BINDING OF THE EASYSEP™ MAGNETIC PARTICLE AFFECT THE CELLS? IS THE FUNCTION OF POSITIVELY SELECTED CELLS ALTERED BY THE BOUND PARTICLES?
      A. Hundreds of publications have used cells selected with EasySep™ positive selection kits for functional studies. Our in-house experiments also confirm that selected cells are not functionally altered by the EasySep™ magnetic particles.

      If particle binding is a key concern, we offer two options for negative selection. The EasySep™ negative selection kits can isolate untouched cells with comparable purities, while RosetteSep™ can isolate untouched cells directly from whole blood without using particles or magnets.

    MODIFYING THE EASYSEP™ PROTOCOL

    • Q. CAN I ALTER THE SEPARATION TIME IN THE MAGNET?
      A. Yes; however, this may impact the kit's performance. The provided EasySep™ protocols have already been optimized to balance purity, recovery and time spent on the isolation.
    • Q. FOR POSITIVE SELECTION, CAN I PERFORM MORE THAN 3 SEPARATIONS TO INCREASE PURITY?
      A. Yes, the purity of targeted cells will increase with additional rounds of separations; however, cell recovery will decrease.

This product has been used in:

  1. Rinath Jeselsohn et al. Cyclin D1 kinase activity is required for the self-renewal of mammary stem and progenitor cells that are targets of MMTV-ErbB2 tumorigenesis.Cancer Cell 17 (1) 65-76 (January 19, 2010)
  2. Norazizah Shafee et al. Cancer stem cells contribute to cisplatin resistance in Brca1/p53-mediated mouse mammary tumors.Cancer Res 68 (9) 3243-3250 (May 1, 2008)
  3. John Stingl et al. Purification and unique properties of mammary epithelial stem cells.Nature advanced online publication (7079) 993-997 (January 4, 2006)

Product Name

Description

Catalog #
Collagenase/Hyaluronidase 10X Collagenase/Hyaluronidase in DMEM 07912
EasySep® Magnet Immunomagnetic Column-Free Magnet 18000
The Big Easy EasySep® Magnet Immunomagnetic Column-Free Magnet 18001
RoboSep™ RoboSep™ - The Fully Automated Cell Separator 20000
EpiCult®-B Mouse Medium Kit Medium for Mouse Mammary Epithelial Cell Culture 05610

FACS Profile Results with EasySep® Mouse Mammary Stem Cell Enrichment Kit


FACS Profile Results with EasySep® Mouse Mammary Stem Cell Enrichment Kit
Flow cytometry profile of freshly dissociated mouse mammary epithelial cells distributed according to their expression of CD49f and CD24. The mammary repopulating unit (MRU)-rich fraction is found in the CD49f++CD24+subset. The mammary colony-forming cells (Ma-CFCs) are found in the CD49f+CD24++ subset.


Typical EasySep® Procedure for Mouse Cells


Typical EasySep® Procedure for Mouse Cells



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