EasySep™ Mouse CD4+ T Cell Enrichment Kit

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Immunomagnetic Negative Selection Kit

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  • Box top label for Mouse CD4+ T Cell Enrichment Kit For labeling 10e9 cells
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The EasySep™ Mouse CD4+ T Cell Enrichment Kit is designed to isolate CD4+ T cells from single cell suspensions of splenocytes or other tissues by negative selection. Unwanted cells are targeted for removal with biotinylated antibodies directed against non-CD4+ T cells (CD8, CD11b, CD11c, CD19, CD45R/B220, CD49b, TER119) and Tetrameric Antibody Complexes recognizing biotin and dextran-coated magnetic particles. Labeled cells are separated using an EasySep™ magnet without the use of columns. Desired cells are poured off into a new tube.
Product Name Description Catalog # Size Price Quantity
EasySep™ Mouse CD4+ T Cell Enrichment Kit Immunomagnetic isolation of untouched mouse CD4+ T cells 19752 For labeling up to 1 x 10e9 cells 539.00 USD      
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RoboSep™ Mouse CD4+ T Cell Enrichment Kit with Filter Tips EasySep™ kit with RoboSep™ buffer and RoboSep™ filter tips 19752RF For labeling up to 1 x 10e9 cells 572.00 USD      
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Call us: +1.604.877.0713  
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Recommended for:
The isolation of mouse CD4+ T cells from single cell suspensions of splenocytes or other tissues by negative selection
Components:
• EasySep™ Mouse CD4+ T Cell Enrichment Cocktail, 0.5 mL
• EasySep™ Biotin Selection Cocktail 2, 1.0 mL
• EasySep™ D2 Magnetic Particles, 1.0 mL
• Normal Rat Serum, 2.0 mL
Intended Use Statement: For Research Use Only. Not for Therapeutic or Diagnostic Use.
Equipment Required:
• EasySep™ Magnet (Catalog #18000), or
• “The Big Easy” EasySep™ Magnet (Catalog #18001), or
• RoboSep™ (Catalog #20000)
Product Type: Cell isolation products
Cell Type: T cells, CD4+ T cells
Popular Product Line: EasySep, RoboSep
Area of Interest: Immunology
Species: Mouse
Cell Source: Spleen, Other
Selection Method: Negative
Cell Isolation Product Type: Reagents

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. Yoshiharu Ohoka et al. Retinoic acid-induced CCR9 expression requires transient TCR stimulation and cooperativity between NFATc2 and the retinoic acid receptor/retinoid X receptor complex.J Immunol 186 (2) 733-744 (January 15, 2011)
  2. J Scott Hale et al. Bcl-2-interacting mediator of cell death influences autoantigen-driven deletion and TCR revision.J Immunol 186 (2) 799-806 (January 15, 2011)
  3. Lori Lebson et al. Cutting edge: The transcription factor Kruppel-like factor 4 regulates the differentiation of Th17 cells independently of ROR??t.J Immunol 185 (12) 7161-7164 (December 15, 2010)
  4. J Scott Hale et al. TCR revision generates functional CD4(+) T cells.J Immunol 185 (11) 6528-6534 (December 1, 2010)
  5. Hajime Takeuchi et al. Efficient induction of CCR9 on T cells requires coactivation of retinoic acid receptors and retinoid X receptors (RXRs): exaggerated T Cell homing to the intestine by RXR activation with organotins.J Immunol 185 (9) 5289-5299 (November 1, 2010)
  6. Shoshana D Katzman et al. Duration of antigen receptor signaling determines T-cell tolerance or activation.Proc Natl Acad Sci U S A 107 (42) 18085-18090 (October 19, 2010)
  7. Sandra Balkow et al. LFA-1 activity state on dendritic cells regulates contact duration with T cells and promotes T-cell priming.Blood 116 (11) 1885-1894 (September 16, 2010)
  8. Denise E De Almeida et al. Immune dysregulation by the rheumatoid arthritis shared epitope.J Immunol 185 (3) 1927-1934 (August 1, 2010)
  9. Carla A Da Silva et al. Chitin Particles are Multifaceted Immune Adjuvants.Am J Respir Crit Care Med (July 23, 2010)
  10. Amanda C Poholek et al. In vivo regulation of Bcl6 and T follicular helper cell development.J Immunol 185 (1) 313-326 (July 1, 2010)
  11. Bernhard Lehnertz et al. Activating and inhibitory functions for the histone lysine methyltransferase G9a in T helper cell differentiation and function.The Journal of Experimental Medicine 207 (5) 915-922 (April 26, 2010)
  12. Jieh-Juen Yu et al. Francisella tularensis T-cell antigen identification using humanized HLA-DR4 transgenic mice.Clin Vaccine Immunol 17 (2) 215-222 (February 2010)
  13. Norbert H??ser et al. Intact LFA-1 deactivation promotes T-cell activation and rejection of cardiac allograft.Int Immunol 22 (1) 35-44 (January 2010)
  14. Sandrine Crab?? et al. The IL-27 p28 subunit binds cytokine-like factor 1 to form a cytokine regulating NK and T cell activities requiring IL-6R for signaling.J Immunol 183 (12) 7692-7702 (December 15, 2009)
  15. Benjamin Marks et al. Thymic self-reactivity selects natural interleukin 17-producing T cells that can regulate peripheral inflammationNat Immunol 10 (10) 1125-1132 (October 2009)
  16. Jonathan H Esensten et al. T-bet-deficient NOD mice are protected from diabetes due to defects in both T cell and innate immune system function.J Immunol 183 (1) 75-82 (July 1, 2009)
  17. Anna Jankowska et al. Loss of heterozygosity 4q24 and TET2 mutations associated with myelodysplastic/myeloproliferative neoplasmsBlood 113 (25) (June 18, 2009)
  18. Moutih Rafei et al. Mesenchymal Stromal Cells Ameliorate Experimental Autoimmune Encephalomyelitis by Inhibiting CD4 Th17 T Cells in a CC Chemokine Ligand 2-Dependent MannerJ Immunol 182 (10) 5994-6002 (May 15, 2009)
  19. Heidi Elsaesser et al. IL-21 Is Required to Control Chronic Viral InfectionScience 324 (5934) 1174182 (May 7, 2009)
  20. Roberto Maldonado et al. Control of T helper cell differentiation through cytokine receptor inclusion in the immunological synapseThe Journal of Experimental Medicine 206 (4) (April 6, 2009)
  21. Moutih Rafei et al. Selective Inhibition of CCR2 Expressing Lymphomyeloid Cells in Experimental Autoimmune Encephalomyelitis by a GM-CSF-MCP1 FusokineJ Immunol 182 (5) 2620-2627 (March 1, 2009)
  22. Yelena Glinka et al. Neuropilin-1 is a receptor for transforming growth factor beta-1, activates its latent form, and promotes regulatory T cell activity.J Leukoc Biol 84 (1) 302-310 (July 2008)
  23. Shunsuke Furuta et al. Overlapping and distinct roles of STAT4 and T-bet in the regulation of T cell differentiation and allergic airway inflammation.J Immunol 180 (10) 6656-6662 (May 15, 2008)
  24. Akira Suto et al. Development and characterization of IL-21-producing CD4+ T cells.J. Exp. Med. 205 (6) 1369-20072057 (May 12, 2008)
  25. Frances Vu et al. ICOS, CD40, and Lymphotoxin {beta} Receptors Signal Sequentially and Interdependently to Initiate a Germinal Center ReactionJ Immunol 180 (4) 2284-2293 (February 15, 2008)
  26. Leslie E Summers-DeLuca et al. Expression of lymphotoxin-alphabeta on antigen-specific T cells is required for DC function.J Exp Med 204 (5) 1071-1081 (May 14, 2007)
  27. Saso Cemerski et al. The stimulatory potency of T cell antigens is influenced by the formation of the immunological synapse.Immunity 26 (3) 345-355 (March 2007)
  28. Heather H Pua et al. A critical role for the autophagy gene Atg5 in T cell survival and proliferation.J Exp Med 204 (1) 25-31 (January 22, 2007)
  29. Wendy E Walker et al. Absence of innate MyD88 signaling promotes inducible allograft acceptance.J Immunol 177 (8) 5307-5316 (October 15, 2006)

Product Name

Description

Catalog #
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

FACS Histogram Results with EasySep® Mouse CD4+ T Cell Enrichment Kit


FACS Histogram Results with EasySep® Mouse CD4+ T Cell Enrichment Kit
Starting with mouse splenocytes, the CD4+ cell content of the enriched fraction typically ranges from 84 - 95%.


Typical EasySep® Procedure for Mouse Cells


Typical EasySep® Procedure for Mouse Cells



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