EasySep™ Release Human CD45 Positive Selection Kit

Immunomagnetic positive selection kit using particle release technology

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Immunomagnetic positive selection kit using particle release technology
From: 661 USD

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Overview

The EasySep™ Release Human CD45 Positive Selection Kit is designed to isolate CD45+ leukocytes from single-cell suspensions of primary human tissues and tumors, or tissues and tumor xenografts from humanized mice, by immunomagnetic positive selection.

Desired cells are labeled with antibodies and magnetic particles, and separated without columns using an EasySep™ magnet. Unwanted cells are simply poured off, while desired cells remain in the tube. Then, bound magnetic particles are removed from the EasySep™-isolated CD45+ cells, which are immediately available for downstream applications. Following cell isolation with this EasySep™ Release kit, antibody complexes remain bound to the cell surface and may interact with Brilliant Violet™ antibody conjugates, polyethylene glycol-modified proteins or other chemically related ligands.
Advantages:
• Highly purified CD45+ cells isolated in less than 45 minutes
• No-wash removal of EasySep™ Releasable RapidSpheres™
Components:
  • EasySep™ Release Human CD45 Positive Selection Kit (Catalog #100-0105)
    • EasySep™ Release Human CD45 Positive Selection Cocktail, 0.5 mL
    • EasySep™ Releasable RapidSpheres™, 1 mL
    • EasySep™ Release Buffer (Concentrate), 6 x 1 mL
  • RoboSep™ Release Human CD45 Positive Selection Kit (Catalog #100-0108)
    • EasySep™ Release Human CD45 Positive Selection Cocktail, 0.5 mL
    • EasySep™ Releasable RapidSpheres™, 1 mL
    • EasySep™ Release Buffer (Concentrate), 6 x 1 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)
    • EasySep™ EasyTube™-14 (Catalog #20128)
  • EasySep™ Release Human CD45 Positive Selection Kit for Humanized Mice (Catalog #100-0107)
    • EasySep™ Release Human CD45 Positive Selection Cocktail, 0.5 mL
    • EasySep™ Releasable RapidSpheres™, 1 mL
    • EasySep™ Release Buffer (Concentrate), 6 x 1 mL
    • Normal Rat Serum, 2 mL
    • EasySep™ Mouse FcR Blocker, 0.5 mL
  • RoboSep™ Release Human CD45 Positive Selection Kit for Humanized Mice (Catalog #100-0109)
    • EasySep™ Release Human CD45 Positive Selection Cocktail, 0.5 mL
    • EasySep™ Releasable RapidSpheres™, 1 mL
    • EasySep™ Release Buffer (Concentrate), 6 x 1 mL
    • Normal Rat Serum, 2 mL
    • EasySep™ Mouse FcR Blocker, 0.5 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125) x 2
    • EasySep™ EasyTube™-14 (Catalog #20128) x 2
Magnet Compatibility:
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• EasyPlate™ EasySep™ Magnet (Catalog #18102)
• RoboSep™-S (Catalog #21000)
Subtype:
Cell Isolation Kits
Cell Type:
Leukocytes
Species:
Human; Mouse
Sample Source:
Other
Selection Method:
Positive
Application:
Cell Isolation
Brand:
EasySep; RoboSep
Area of Interest:
Cancer Research; Immunology

Scientific Resources

Product Documentation

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Educational Materials

(6)

Data and Publications

Data

Using the EasySep™ Release Human CD45 Positive Selection Kit, the CD45+ TIL purities of the start and final isolated fractions are 5.5% and 96.0%, respectively.

Figure 1. EasySep™ Release Human CD45 Positive Selection Kit

Starting with a single-cell suspension of a human breast cancer tumor xenograft (MDA-MB-231) sample from an NRG-3GS humanized mouse, the CD45+ TIL purities of the start and final isolated fractions are 5.5% and 96.0%, respectively.
NOTE: Cell debris and dead cells were excluded from the analysis based on DRAQ5™ and DAPI fluorescence.

Using PBMCs as a startnig sample, this t-SNE plot shows that EasySep™-isolated CD45+ cells are representative of the starting leukocyte population.

Figure 2. EasySep™-Isolated CD45+ Cells are Representative of the Starting Leukocyte Population

Mass cytometry data comparing the composition of immune subsets in PBMCs and EasySep™-isolated cells from the same donor. Starting with whole blood, PBMCs were prepared by density gradient centrifugation using Lymphoprep™. To compare immune subset composition pre- and post-EasySep™ isolation, a fraction of the PBMCs was further isolated using EasySep™ Release Human CD45 Positive Selection Kit and the pre- and post-isolated fractions were assessed by mass cytometry (CyTOF®). t-SNE plots of cells stained with 19 markers and analyzed by CyTOF® are shown (n = 1).

PBMCs pre- and post-EasySep™ isolation were incubated in peptide pools (CEF, CPI, or PHA) and a robust population of IFN-gamma-producing cells in these respective samples are visible on ELISpot plates, showing that EasySep™-isolated CD45+ cells produce IFN-gamma in response to antigen and mitogen stimulation.

Figure 3. EasySep™-Isolated CD45+ Cells Produce IFN-gamma in Response to Antigen and Mitogen Stimulation

PBMCs pre- and post-EasySep™ isolation were incubated for 24 hours in the presence of peptide pools (CEF for antigen-specific CD8+ T cell response and CPI for antigen-specific CD4+ T cell response) or mitogen (phytohemagglutinin [PHA]). Following incubation, ELISpot plates were processed and IFN-gamma-producing cells were counted using an AID ELISpot reader. Representative images of ELISpot assays are shown (n = 3).

CD45+ cells isolated from spleens, lungs, bone marrow, MDA MB231 tumors, and SKOV3 tumors using EasySep™ are highly purified.

Figure 4. CD45+ Cells Isolated by EasySep™ from Various Tissues Are Highly Purified

A humanized mouse tumor model was generated by engraftment of human CD34+ hematopoietic stem and progenitor cells into NRG-3GS mice followed by xenotransplantation with human cancer cell lines, MDA-MB-231 (breast cancer) or SKOV-3 (ovarian carcinoma). Starting with a single-cell suspension of spleens, lungs, bone marrow, or tumors, human CD45+ leukocytes were isolated using EasySep™ Release Human CD45 Positive Selection Kit. The starting frequencies and isolated purities of human CD45+ cells of individual experiments and averages are shown.

PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.