StemSep™ Human Monocyte Enrichment Kit

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Immunomagnetic Column-Based Negative Selection Kit

Free from STEMCELL

  • StemSep™ Box
  • Box top label for Human Monocyte Enrichment Kit for labeling 1x10e9 cells
  • Box top label for Human Monocyte Starter Kit For labeling 10e9 cells.
StemSep™ Box
The StemSep™ Human Monocyte Enrichment Kit is designed to isolate monocytes from a peripheral blood mononuclear cell suspension by negative selection. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing CD2, CD3, CD19, CD56, CD66b, glycophorin A and dextran-coated magnetic particles. Labeled cells are separated using StemSep™ columns and magnets.
Product Name Description Catalog # Size Price Quantity
StemSep™ Human Monocyte Enrichment Kit Column-based Immunomagnetic negative selection of human monocytes 14058 For labeling up to 1 x 10e9 cells 745.00 USD      
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StemSep™ Human Monocyte Enrichment Kit Column-based Immunomagnetic negative selection of human monocytes 14068 For labeling up to 5 x 10e9 cells 3045.00 USD      
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Product Type: Cell isolation products
Recommended For:
The isolation of human monocytes from a peripheral blood mononuclear cell suspension by negative selection
Components:
• StemSep™ Human Monocyte Enrichment Cocktail, 2.0 mL (5 x 2.0 mL)
• StemSep™ Magnetic Colloid, 1.5 mL (4 x 1.5 mL)
Equipment Required:
• StemSep™ Magnet (Catalog #11030, 11050, 11060, or 11070) or a magnet with the strength of at least 0.5 Tesla
• StemSep™ Negative Selection Columns
Cell Type: Monocytes
Popular Product Line: StemSep™
Area of Interest: Immunology
Species: Human
Cell Source: PBMC
Selection Method: Negative
Cell Isolation Product Type: Reagents
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

Procedures and instruction manuals:

MSDS:

FAQS:


  • STEMSEP™ PROCEDURE

    • Q. WHAT CONCENTRATION SHOULD CELLS BE SUSPENDED AT PRIOR TO SEPARATION?
      A. A cell concentration of 2-8 x 107 is acceptable. If using 1.0" columns with human cells, the cell concentration is increased to 108 cells/mL.
    • Q. WHAT IS THE MINIMUM SAMPLE SIZE THAT CAN BE USED?
      A. The minimum volume in which cells should be suspended is 100 µL for human cell separations and 200 µL for mouse cell separations. The volume of cell suspension determines the volume of cocktail and colloid to use. If small numbers of cells are to be separated, the suspension may be less concentrated than 2x107 cells/mL.
    • Q. WHAT IS THE BENEFIT OF USING A PUMP?
      A. The benefit is twofold: 1) It can allow up to four simultaneous separations to be performed when used in conjunction with the blue magnet (#11060). 2) It can improve purity and recovery when larger columns are used.
    • Q. SHOULD MY PRIMING BUFFER CONTAIN PROTEIN?
      A. The priming medium should not contain protein. Use PBS, either at room temperature or degassed, without serum or other protein to prime the column from bottom up. However, the separation medium used to wash the column prior to use will contain protein.
    • Q. DO BUFFERS NEED TO BE DEGASSED PRIOR TO DOING STEMSEP™?
      A. No. Degassing the buffer is not necessary.
    • Q. WHAT IS THE FUNCTION OF EDTA IN THE BUFFER?
      A. EDTA prevents cells such as monocytes from adhering to the column. The effective concentration is 1–2 mM.
    • Q. WHY IN THE STEMSEP™ MANUAL IS IT SUGGESTED TO AVOID DEXTRAN SEDIMENTATION?
      A. Our particles are coated with dextran, so if the free dextran is not washed away completely there could be some interference with colloid binding. HetaSep sedimentation is a good alternative to dextran. Please see the HetaSep protocol.
    • Q. CAN THE STEMSEP™ COLUMNS BE REUSED?
      A. Yes, we have found that the columns can be reused up to six times without a decrease in performance, see protocol for Reusing StemSep Columns.
    • Q. CAN THE CELLS THAT WERE RETAINED IN THE COLUMN BE REMOVED?
      No. Since our columns bind labeled cells so strongly, it is difficult to remove cells from the column even after the column is removed from the magnet.
    • Q. WHY IS IT SUGGESTED THAT THE ANTI-CD32 ANTIBODY BE USED WHEN ISOLATING MONOCYTES?
      A. Monocytes may non-specifically bind tetramers via the Fcg RIIA on their surface, and subsequently be retained on the column. To decrease this non-specific binding, cells should be incubated with an anti-CD32 blocking reagent (Catalog #14531, #14551, and #14561) prior to immunomagnetic labeling. To use, follow the instructions in the StemSep™ Operating Manual.

    STEMSEP™ COMPABILITY

    • Q. CAN STEMSEP™ COLUMNS BE USED FOR POSITIVE SELECTION?
      A. No. Our positive selection reagents can be used with commercially available positive selection columns.
    • Q. CAN I USE STEMSEP™ ON REAGENTS OTHER MAGNETIC CELL SEPARATION SYSTEMS?
      A. Because of the extremely small size of the StemSep™ magnetic colloid (50-150 nm), we have found that StemSepTM reagents are completely compatible with any magnet of strength equal or greater than 0.5 Tesla. Many customers have reported that their results have been equivalent or better with the economical StemSep™ reagents.
    • Q. WHICH SPECIES CAN STEMSEP™ BE USED WITH?
      A. Cocktails have been optimized for the selection of human, mouse, rat and non-human primate samples.

    CLUMPY SAMPLES

    • Q. ARE THERE ANY SUGGESTIONS FOR CLUMPY SAMPLES?
      A. Yes. There are several reasons a sample might clump.
      1) If the cells were frozen or poorly stored then DNA from dead cells can form clumps. If this is the case adding DNAse to the cell suspension may help.
      2) The presence of Ca++/Mg++ in the medium may cause cells, such as monocytes, to clump. Ca++/Mg++ free PBS should be used. EDTA can also be used to decrease clumping.
      3) Cells may clump if they become activated. Keeping the cells at 4°C or on ice can prevent this.
    • Q. MY CORD BLOOD SAMPLE IS CLUMPING. WHAT IS HAPPENING?
      A. Cord blood can frequently be “clumpy”. Often, this is a platelet problem, so the slow spin procedure outlined in the StemSep™ Instruction Manual (pg. 33) should be followed.
    • Q. WHICH ANTICOAGULANT SHOULD BE USED?
      A. ACD is better than heparin as it is less likely to give a clumpy or sticky cell suspension.

This product has been used in:

  1. Nicholas M Stamatos et al. LPS-induced cytokine production in human dendritic cells is regulated by sialidase activity.J Leukoc Biol (September 8, 2010)

Product Name

Description

Catalog #

CD14 Antibody, Clone MoP9, FITC-Conjugated Mouse Monoclonal Antibody to Human CD14 - FITC Conjugated 10406
4 Channel Peristaltic Pump StemSep™ Equipment 11000
StemSep™ Red Magnet with Stand Immunomagnetic Column-Based Magnet 11030
StemSep™ Green Magnet with Stand Immunomagnetic Column-Based Magnet 11050
StemSep™ Blue Magnet with Stand
Immunomagnetic Column-Based Magnet
11060
StemSep™ Black Magnet with Stand Immunomagnetic Column-Based Magnet 11070

Typical StemSep™ Monocyte Enrichment Profile


FACs Plot
Starting with a previously frozen peripheral blood mononuclear cell suspension, the CD14+ cell content of the enriched fraction typically ranges from 80 - 89%.


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