EasySep™ Mouse CD8+ T Cell Enrichment Kit

Procedures and instruction manuals:

•  PRODUCT INFORMATION SHEET - 19753, Lot# 11H40541 or less:28753-PIS_3_2_0.pdf
•  PRODUCT INFORMATION SHEET - 19753, Lot# 11H40542 or greater:28753-PIS_4_0_0.pdf
•  PRODUCT INFORMATION SHEET - 19753, Lot# 11H40542 or greater:28753-EasyPlate-PIS_1_0_0.pdf
•  PRODUCT INFORMATION SHEET - 19753RF, Lot# 11H40541 or less:28753-PIS_3_2_0.pdf
•  PRODUCT INFORMATION SHEET - 19753RF, Lot# 11H40542 or greater:28753-PIS_4_0_0.pdf

Educational resources:

•  VIDEO: EasySep®: Powerful Immunomagnetic Isolation of Virtually Any Cell Type
•  VIDEO: The EasySep® Pour-off: A Simple and Effective Way to Isolate Cells
•  VIDEO: EasySep®: Powerful Immunomagnetic Isolation of Virtually Any Cell Type
•  BROCHURE: Cell Separation Equipment & Products
•  BROCHURE: Cell Separation for Autoimmunity Research
•  WALLCHART: Antigen Processing and Presentation Wallchart
•  BROCHURE: Ready Sep Go – Cell Separation In As Little As 25 Minutes
•  VIDEO: The EasySep® MultiStand: Four Samples, Four Separations, One Pour-off
•  BROCHURE: Customized Cell Isolation: Flexible Cell Isolation Designed for you and your Research
•  VIDEO: HetaSep™: A Guide to Isolating Nucleated Cells from Whole Blood
•  VIDEO: RoboSep® Product Tour
•  VIDEO: Ficoll-Paque™ PLUS: A Guide to Isolating Mononuclear Cells from Whole Blood
•  VIDEO: RoboSep® Sneak Preview

MSDS:

•  19713C.4_19753C.4-MSDS.pdf
•  19150.1-MSDS.pdf

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. Maciej Kujawski et al. Targeting STAT3 in adoptively transferred T cells promotes their in vivo expansion and antitumor effects.Cancer Res 70 (23) 9599-9610 (December 1, 2010)
2. Andreas Herrmann et al. Targeting Stat3 in the myeloid compartment drastically improves the in vivo antitumor functions of adoptively transferred T cells.Cancer Res 70 (19) 7455-7464 (October 1, 2010)
3. 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)
4. 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)
5. Christian S Hinrichs et al. Adoptively transferred effector cells derived from naive rather than central memory CD8+ T cells mediate superior antitumor immunity.Proc Natl Acad Sci U S A 106 (41) 17469-17474 (October 13, 2009)
6. Mo??ra Fran??ois et al. Mesenchymal stromal cells cross-present soluble exogenous antigens as part of their antigen-presenting cell properties.Blood 114 (13) 2632-2638 (September 24, 2009)
7. Christopher M. Snyder et al. CD4+ T Cell Help Has an Epitope-Dependent Impact on CD8+ T Cell Memory Inflation during Murine Cytomegalovirus InfectionJ Immunol 183 (6) 3932-3941 (September 15, 2009)
8. Gabrielle Rizzuto et al. Self-antigen???specific CD8+ T cell precursor frequency determines the quality of the antitumor immune responseThe Journal of Experimental Medicine 206 (4) (April 13, 2009)
9. Lydia Makaroff et al. Postthymic maturation influences the CD8 T cell response to antigenProceedings of the National Academy of Sciences of the United States of America 106 (12) 4799-4804 (March 24, 2009)
10. 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)
11. Christopher Snyder et al. Immunity 29 (4) 650-659 (October 17, 2008)
12. 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)
13. 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)
14. Wendy E Walker et al. Absence of innate MyD88 signaling promotes inducible allograft acceptance.J Immunol 177 (8) 5307-5316 (October 15, 2006)

FACS Histogram Results with EasySep® Mouse CD8+ T Cell Enrichment Kit

Typical EasySep® Procedure for Mouse Cells