RosetteSep™ HLA Myeloid Cell Enrichment Kit

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Immunodensity Negative Selection Cocktail



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  • RosetteSep® Box
  • Box top label for RosetteSep® HLA Human Myeloid Cell Enrichment Kit for Chimerism Analysis
RosetteSep® Box
The RosetteSep™ HLA Myeloid Cell Enrichment Cocktail is designed to enrich myeloid (CD33+) cells from whole blood by negative selection for lineage-specific chimerism analysis. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing CD3, CD8, CD19, CD56 and glycophorin A on red blood cells (RBCs). When centrifuged over a buoyant density medium such as RosetteSep™ DM-M (Catalog #15725), the unwanted cells pellet along with the RBCs. The purified myeloid cells are present as a highly enriched population at the interface between the plasma and the buoyant density medium. The desired cells are immediately ready for chimerism analysis.
 
This product has been CE Marked in the European Union according to the 98/79/EC In Vitro Diagnostic Device Directive and complies with its requirements.
Product Name Description Catalog # Size Price Quantity
RosetteSep™ HLA Myeloid Cell Enrichment Kit Immunodensity isolation of untouched myeloid cells for lineage-specific chimerism analysis 15272HLA For 20 tests (10 mL of whole blood per test) 782.00 USD      
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Recommended for:
The isolation of human myeloid cells from whole blood by negative selection for HLA analysis
Components:
• RosetteSep™HLA Myeloid Cell Enrichment Cocktail, 5 x 2.0 mL
• RosetteSep™ DM-M, 2 x 100 mL
Legal Statement: Ficoll-Paque™ PLUS is a trademark of GE Healthcare Ltd.
Product Type: Cell isolation products
Area of Interest: Chimerism, HLA
Cell Isolation Product Type: Reagents
Cell Source: Buffy coat, Whole blood
Cell Type: Myeloid cells
Popular Product Line: RosetteSep
Selection Method: Negative
Species: Human

Procedures and instruction manuals:

Educational resources:

MSDS:

FAQS:


  • HOW ROSETTESEP™ WORKS

    • 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 density centrifugation medium (e.g. Ficoll-Paque™ PLUS, Lymphoprep™).
    • Q. WHAT FACTORS AFFECT CELL RECOVERY?
      A. The temperature of the reagents can affect cell recovery. All reagents should be at room temperature (sample, density centrifugation medium, PBS, centrifuge) before performing the isolations. Layering can also affect recovery so be sure to carefully layer the sample to avoid mixing with the density centrifugation medium as much as possible. Be sure to collect the entire enriched culture without disturbing the RBC pellet. A small amount of density centrifugation medium can be collected without worry.

    CELL TYPES AND SAMPLE SOURCES

    • Q. WHICH CELL SAMPLES CAN ROSETTESEP™ BE USED WITH?
      A. RosetteSep™ can be used with leukapheresis samples, bone marrow 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).
    • 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. 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™, 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 cell isolation system which works with mouse and other non-human species.

    ANTI-COAGULANTS

    • Q. WHICH ANTICOAGULANT SHOULD BE USED WITH ROSETTESEP™?
      A. Peripheral blood should be collected in heparinized Vacutainers. 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.

Product Name

Description

Catalog #

RosetteSep™ DM-M Density Medium Density Gradient Medium 15725

FACS Histogram Results with RosetteSep® Human Myeloid Cell Enrichment Kit


FACS Histogram Results with RosetteSep® Human Myeloid Cell Enrichment Kit