RosetteSep® Human CD3 Depletion Cocktail

Procedures and instruction manuals:

•  PRODUCT INFORMATION SHEET - 15621, Lot# All:28585-PIS_1_0_0.pdf
•  PRODUCT INFORMATION SHEET - 15661, Lot# All:28585-PIS_1_0_0.pdf

Educational resources:

•  VIDEO: RosetteSep® Introduction
•  VIDEO: RosetteSep®: A Guide to Isolating NK Cells Directly from Whole Blood
•  BROCHURE: One-Step Cell Enrichment Directly From Whole Blood
•  BROCHURE: Cell Separation for Autoimmunity Research
•  BROCHURE: Ready Sep Go – Cell Separation In As Little As 25 Minutes
•  VIDEO: RosetteSep® for HLA Analysis

MSDS:

•  15611C_15621C-MSDS.pdf

FAQS:

• 
  Q. WHAT IS ROSETTESEP^®?
  
  A. RosetteSep^® is a rapid cell separation procedure for the isolation of purified cells directly from whole blood without columns or magnets.
• 
  Q. HOW DOES ROSETTESEP^® WORK?
  
  A. The antibody cocktail crosslinks unwanted cells to red blood cells (RBCs), forming rosettes. The unwanted cells then pellet with the free RBCs when centrifuged over a buoyant density medium.
• 
  Q. WHICH CELL SAMPLES CAN ROSETTESEP^® BE USED WITH?
  
  A. RosetteSep^® can be used with leukapheres, bone marrow samples or buffy coat, as long as: the concentration of cells does not exceed 5 x 10⁷ per mL (can dilute if necessary); and there are at least 20 RBCs for every nucleated cell (RBCs can be added if necessary).  For a full procedure click here.
• 
  Q. CAN ROSETTESEP^® BE USED WITH PREVIOUSLY FROZEN OR CULTURED CELLS?
  
  A. Yes. Cells should be re-suspended at 2 – 5 x 10⁷ cells / mL in PBS + 2% FBS. Fresh whole blood should be added at 250 µL per mL of sample, as a source of red cells.
• 
  Q. WHICH ANTICOAGULANT SHOULD BE USED WITH ROSETTESEP^®?
  
  A. Peripheral blood should be collected in heparinized Vaccutainers. Cord blood should be collected in ACD.
• 
  Q. SHOULD THE ANTICOAGULANT BE WASHED OFF BEFORE USING ROSETTESEP^®?
  
  A. No, the antibody cocktail can be added directly to the sample.
• 
  Q. CAN ROSETTESEP^® BE USED TO ENRICH PROGENITORS FROM CORD BLOOD?
  
  A. Yes. Sometimes cord blood contains immature nucleated red cells that have a lower density than mature RBCs. These immature red cells do not pellet over Ficoll^TM, which can lead to a higher RBC contamination than peripheral blood separations.
• 
  Q. DOES ROSETTESEP^® WORK WITH MOUSE CELLS?
  
  A. No, but we have developed EasySep^®, a magnetic-based separation for the isolation of mouse cells.

This product has been used in:

1. Todd M Brusko et al. Human antigen-specific regulatory T cells generated by T cell receptor gene transfer.PLoS One 5 (7) e11726 (2010)
2. Weihong Liu et al. CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4(+) T reg cells.J Exp Med 203 (7) 1701-1711 (July 10, 2006)
3. Konstantina Nika et al. Lipid raft targeting of hematopoietic protein tyrosine phosphatase by protein kinase C theta-mediated phosphorylation.Mol Cell Biol 26 (5) 1806-1816 (March 2006)
4. Xiao-Song He et al. T cell-dependent production of IFN-gamma by NK cells in response to influenza A virus.J Clin Invest 114 (12) 1812-1819 (December 2004)

FACS Histogram Results Using RosetteSep® Human CD3+ Cell Depletion Cocktail

RosetteSep® Procedure