RosetteSep™ Human Monocyte Enrichment Cocktail

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Immunodensity Negative Selection Cocktail

Free from STEMCELL

  • RosetteSep™ Box
  • Box top label for RosetteSep™ Human Monocyte Enrichment Cocktail For labeling 40 mL blood
  • Box top label for RosetteSep™ Human Monocyte Enrichment Cocktail For labeling 200 mL blood
RosetteSep™ Box
The RosetteSep™ Human Monocyte Enrichment Cocktail is designed to isolate monocytes from whole blood by negative selection. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes (TAC) recognizing CD2, CD3, CD8, CD19, CD56, CD66b, CD123 and glycophorin A on red blood cells (RBCs). When centrifuged over a buoyant density medium such as Lymphoprep™ (Catalog #07801), the unwanted cells pellet along with the RBCs. The purified monocytes are present as a highly enriched population at the interface between the plasma and the buoyant density medium.
Product Name Description Catalog # Size Price Quantity
RosetteSep™ Human Monocyte Enrichment Cocktail Immunodensity isolation of untouched monocytes 15068 For labeling 200 mL of whole blood 599.00 USD      
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RosetteSep™ Human Monocyte Enrichment Cocktail Immunodensity isolation of untouched monocytes 15028 For labeling 40 mL of whole blood 155.00 USD      
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Product Type: Cell isolation products
Recommended For:
The isolation of human monocytes from whole blood by negative selection
Components:
RosetteSep™ Human Monocyte Enrichment Cocktail
Accessory Products: Lymphoprep™ (Catalog #07801)
Cell Type: Monocytes
Popular Product Line: RosetteSep™
Area of Interest: Immunology, HIV
Species: Human
Cell Source: Buffy coat, Whole blood
Selection Method: Negative
Cell Isolation Product Type: Reagents
Legal Statement: Ficoll-Paque™ PLUS is a trademark of GE Healthcare Ltd.
Intended Use Statement: FOR RESEARCH USE ONLY. NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES.
ISO Statement: STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485 MEDICAL DEVICE STANDARDS.

Protocols and Manuals

Current documents
Product NameLot# ?Document TypeDownload
RosetteSep™ Human Monocyte Enrichment Cocktail All Product Information Sheet
 

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.
 

This product has been used in:

  1. Sandra Armengol Lopez et al. The oxidative state of chylomicron remnants influences their modulation of human monocyte activation.Int J Vasc Med 2012 942512 (2012)
  2. C Bentley et al. Influence of chylomicron remnants on human monocyte activation in vitro.Nutr Metab Cardiovasc Dis 21 (11) 871-878 (November 2011)
  3. Christian Smedman et al. FluoroSpot analysis of TLR-activated monocytes reveals several distinct cytokine secreting subpopulations.Scand J Immunol (September 28, 2011)
  4. Shinji Okano et al. Provision of continuous maturation signaling to dendritic cells by RIG-I-stimulating cytosolic RNA synthesis of Sendai virus.J Immunol 186 (3) 1828-1839 (February 1, 2011)
  5. Natalia Ferraz et al. Nanoporosity of alumina surfaces induces different patterns of activation in adhering monocytes/macrophages.Int J Biomater 2010 402715 (2010)
  6. A trans-acting locus regulates an anti-viral expression network and type 1 diabetes risk.Nature 467 (7314) 460 (September 8, 2010)
  7. Molly A Ingersoll et al. Comparison of gene expression profiles between human and mouse monocyte subsets.Blood 115 (3) e10-9 (January 21, 2010)
  8. Francisco J. Maldonado-Arocho et al. Anthrax Edema Toxin Induces Maturation of Dendritic Cells and Enhances Chemotaxis towards Macrophage Inflammatory Protein 3{beta}Infect. Immun. 77 (5) 2036-2042 (May 1, 2009)
  9. Jeremy Mauldin et al. Reduced expression of ATP-binding cassette transporter G1 increases cholesterol accumulation in macrophages of patients with type 2 diabetes mellitus.Circulation 117 (21) 2785-2792 (May 27, 2008)
  10. Yumiko Wada et al. Selective abrogation of Th1 response by STA-5326, a potent IL-12/IL-23 inhibitor.Blood 109 (3) 1156-1164 (February 1, 2007)
  11. Nichole L. Korpi-Steiner et al. Human rhinovirus induces robust IP-10 release by monocytic cells, which is independent of viral replication but linked to type I interferon receptor ligation and STAT1 activation.J Leukoc Biol 80 (6) 1364-1374 (December 1, 2006)
  12. Krzysztof Laudanski et al. Cell-specific expression and pathway analyses reveal alterations in trauma-related human T cell and monocyte pathways.Proc Natl Acad Sci U S A 103 (42) 15564-15569 (October 17, 2006)
  13. Jiangning Li et al. cDNA microarray analysis reveals fundamental differences in the expression profiles of primary human monocytes, monocyte-derived macrophages, and alveolar macrophages.J Leukoc Biol 81 (1) 328-335 (October 17, 2006)
  14. Jennifer A Fulcher et al. Galectin-1-matured human monocyte-derived dendritic cells have enhanced migration through extracellular matrix.J Immunol 177 (1) 216-226 (July 1, 2006)
  15. Eva M. Carmona et al. Pneumocystis Cell Wall beta-Glucans Induce Dendritic Cell Costimulatory Molecule Expression and Inflammatory Activation through a Fas-Fas Ligand MechanismJ Immunol 177 (1) 459-467 (July 1, 2006)
  16. Shigeru Tsuboi et al. A Complex of Wiskott-Aldrich Syndrome Protein with Mammalian Verprolins Plays an Important Role in Monocyte ChemotaxisJ Immunol 176 (11) 6576-6585 (June 1, 2006)
  17. Jun Abe et al. Gene Expression Profiling of the Effect of High-Dose Intravenous Ig in Patients with Kawasaki DiseaseJ Immunol 174 (9) 5837-5845 (May 1, 2005)
  18. J Perren Cobb et al. Application of genome-wide expression analysis to human health and disease.Proc Natl Acad Sci U S A 102 (13) 4801-4806 (March 29, 2005)
  19. Francine Garnache-Ottou et al. Expression of the myeloid-associated marker CD33 is not an exclusive factor for leukemic plasmacytoid dendritic cells.Blood 105 (3) 1256-1264 (February 1, 2005)
  20. Ari Rouhiainen et al. Regulation of monocyte migration by amphoterin (HMGB1).Blood 104 (4) 1174-1182 (August 15, 2004)
  21. Fr??d??ric Ebstein et al. Cytotoxic T cell responses against mesothelioma by apoptotic cell-pulsed dendritic cells.Am J Respir Crit Care Med 169 (12) 1322-1330 (June 15, 2004)
  22. Udaykumar Ranga et al. Tat Protein of Human Immunodeficiency Virus Type 1 Subtype C Strains Is a Defective ChemokineJ. Virol. 78 (5) 2586-2590 (March 1, 2004)
  23. Melanie Wellington et al. Enhanced Phagocytosis of Candida Species Mediated by Opsonization with a Recombinant Human Antibody Single-Chain Variable FragmentInfect. Immun. 71 (12) 7228-7231 (December 1, 2003)
  24. James J Kohler et al. Human immunodeficiency virus type 1 (HIV-1) induces activation of multiple STATs in CD4+ cells of lymphocyte or monocyte/macrophage lineages.J Leukoc Biol 73 (3) 407-416 (March 2003)
  25. Claire E Hirst et al. The intracellular granzyme B inhibitor, proteinase inhibitor 9, is up-regulated during accessory cell maturation and effector cell degranulation, and its overexpression enhances CTL potency.J Immunol 170 (2) 805-815 (January 15, 2003)
  26. Peter A Sieling et al. Toll-like receptor 2 ligands as adjuvants for human Th1 responses.J Immunol 170 (1) 194-200 (January 1, 2003)
  27. Bouchentouf M et al. Monocyte Derivatives Promote Angiogensis and Myocyte Survival in a Model of Myocardial Infarction. Cell TransplantationCell Transplantation (epub ahead of print)

Product Name

Description

Catalog #

CD14 Antibody, Clone MoP9, FITC-Conjugated Mouse Monoclonal Antibody to Human CD14 - FITC Conjugated 10406
Lymphoprep™ Density Gradient Medium for the Isolation of Mononuclear Cells 07801

FACS Histogram Results Using RosetteSep™ Human Monocyte Enrichment Cocktail


FACS Histogram Results Using RosetteSep™ Human Monocyte Enrichment Cocktail
Starting with fresh peripheral blood, the CD14+ cell content of the enriched fraction is typically 72% - 85%. *Note: Red blood cells were removed by lysis prior to flow cytometry.


RosetteSep™ Procedure


RosetteSep™ Procedure



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