EasySep™ Direct Human PBMC Isolation Kit

Immunomagnetic negative selection

New look, same high quality and support! You may notice that your instrument or reagent packaging looks slightly different from images displayed on the website, or from previous orders. We are updating our look but rest assured, the products themselves and how you should use them have not changed. Learn more

EasySep™ Direct Human PBMC Isolation Kit

Immunomagnetic negative selection

From: 535 USD
Catalog #
19654_C
Immunomagnetic negative selection

Product Advantages

• 99.9% RBC depletion without the need for density gradient centrifugation, sedimentation or lysis
• Fast, easy-to-use and column-free
• Isolated cells are untouched

What's Included

  • EasySep™ Direct Human PBMC Isolation Kit (Catalog #19654)
    • EasySep™ Direct Human PBMC Isolation Cocktail, 2 x 2.5 mL
    • EasySep™ Direct RapidSpheres™, 4 x 2.5 mL
  • EasySep™ Direct Human PBMC Isolation Kit for RoboSep™ (Catalog #19654RF)
    • EasySep™ Direct Human PBMC Isolation Cocktail, 2 x 2.5 mL
    • EasySep™ Direct RapidSpheres™, 4 x 2.5 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)
New look, same high quality and support! You may notice that your instrument or reagent packaging looks slightly different from images displayed on the website, or from previous orders. We are updating our look but rest assured, the products themselves and how you should use them have not changed. Learn more

Overview

The EasySep™ Direct Human PBMC Isolation Kit is designed to isolate highly purified peripheral blood mononuclear cells (PBMCs) from fresh whole blood, buffy coat, bone marrow, cord blood, or leukapheresis products by negative selection. Red blood cells (RBCs), platelets, and unwanted cells are targeted for removal with antibody complexes and magnetic particles and separated using an EasySep™ magnet. Untouched PBMCs are simply collected into a new tube and are immediately available for downstream applications such as flow cytometry, culture, or DNA/RNA extraction.
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• Easy 50 EasySep™ Magnet (Catalog #18002)
Subtype
Cell Isolation Kits
Cell Type
B Cells, Lymphocytes, Monocytes, Mononuclear Cells, NK Cells, T Cells
Species
Human
Sample Source
Bone Marrow, Buffy Coat, Cord Blood, Leukapheresis, Whole Blood
Selection Method
Negative
Brand
EasySep, RoboSep
Area of Interest
Immunology

Scientific Resources

Product Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
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19654
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All
Language
English
Catalog #
19654
Lot #
All
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English
Catalog #
19654
Lot #
All
Language
English
Catalog #
19654
Lot #
All
Language
English
Catalog #
19654
Lot #
All
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English
Catalog #
19654
Lot #
All
Language
English
Catalog #
19654RF
Lot #
All
Language
English
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19654
Lot #
All
Language
English
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Safety Data Sheet 2
Catalog #
19654
Lot #
All
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English
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Safety Data Sheet 3
Catalog #
19654
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
19654RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
19654RF
Lot #
All
Language
English

Data and Publications

Figure 1. Typical EasySep™ Direct Human PBMC Isolation Profile

In the above example, the mononuclear cell content of the whole blood start sample (lysed by ammonium chloride) and non-lysed final isolated fraction is 27.0% and 98.6% (not gated on CD45), respectively.

Figure 2. Representative Flow Cytometry Plots

Representative FSC vs. SSC flow cytometry plots of mononuclear cells isolated from whole blood samples using either density gradient centrifugation or EasySep™ Direct Human PBMC Isolation Kit.

Figure 3. Representative t-SNE plots of Isolated PBMCs

Representative t-SNE plots of PBMCs stained with 19 markers and analyzed with mass cytometry (CyTOF). Cells are clustered and colored based on the combination of markers they express. Both density gradient centrifugation and EasySep™ Direct Human PBMC Isolation Kit resulted in comparable cell populations with the exception of the contaminating granulocyte population (CD16+CD15+CD11b+CD66b+ CD45low).

Figure 4. EasySep™ Direct Human PBMC Isolation Kit Results in Fewer Contaminating Cells Compared to Density Gradient Centrifugation

Mononuclear cells were isolated from whole blood samples using either density gradient centrifugation or EasySep™ Direct Human PBMC Isolation Kit. Cells were counted and analyzed by flow cytometry. (A) Both density gradient centrifugation and EasySep™ Direct Human PBMC Isolation Kit resulted in an equivalent total number of nucleated cells recovered from 24-hour blood samples (mean ± SD; n=14). (B) Using EasySep™ Direct Human PBMC Isolation Kit to obtain mononuclear cells from 24-hour old blood samples resulted in lower numbers of residual platelets (CD41+), red blood cells (Glycophorin A+/CD45-), and granulocytes (CD66b+) compared to density gradient centrifugation (mean ± SD; n=15). (C) Cell isolation from 24-hour, 48-hour, 72-hour and 96-hour old blood samples using EasySep™ Direct Human PBMC Isolation Kit resulted in lower numbers of residual granulocytes compared to cell isolation using density gradient centrifugation (mean ± SD; n=3).

Figure 5. PBMCs Isolated with EasySep™ Direct Human PBMC Isolation Kit Proliferate and Maintain High RNA Integrity

Mononuclear cells were isolated from whole blood samples using either density gradient centrifugation or EasySep™ Direct Human PBMC Isolation Kit. (A) Isolated mononuclear cells were used for downstream RNA isolation. There was no significant difference in RNA integrity as measured with the Agilent RNA Bioanalyzer (mean ± SEM, n=3). (B) Isolated mononuclear cells were labeled with Proliferation Dye eFluor 450 and stimulated with ImmunoCult™ Human CD3/CD28 T Cell Activator and 0.5ng/ml IL-2. After 4 days in culture, cells were analyzed for proliferation by flow cytometry. Representative histogram showing dividing cells (eFluor 450low).

Figure 6. PBMC Isolation from a Full-Size Leukapheresis Pack Using the EasySep™ Direct Human PBMC Isolation Kit Automated with RoboSep™-S Results in Fewer Contaminating Cells Compared to Density Gradient Centrifugation

Mononuclear cells were isolated from single concentrated leukapheresis packs using either density gradient centrifugation with Lymphoprep™ density gradient medium (Density Gradient Centrifugation) or EasySep™ Direct Human PBMC Isolation Kit automated on the RoboSep™-S instrument (EasySep™ Direct on RoboSep™-S). Cells were counted and analyzed by flow cytometry. Compared to density gradient centrifugation, EasySep™ Direct Human PBMC Isolation Kit automated on the RoboSep™-S instrument resulted in (A) equivalent numbers of total mononuclear cells, (B) equivalent numbers of total monocytes and lymphocytes and (C) lower numbers of residual platelets (CD41+), red blood cells (Glycophorin A+ /CD45− ), and granulocytes (CD66b+ ) (mean ± SD n=6).

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