Simplify Your PBMC Isolations with the EasySep™ Direct Human PBMC Isolation Kit
- Document # 27277
- Version 1.0.0
- Aug 2025
Peripheral blood mononuclear cells (PBMCs) are a cornerstone of immunology, cancer, and infectious disease research. These versatile immune cells, composed of lymphocytes, monocytes, and dendritic cells, enable laboratories to study immune responses, develop novel therapies, and advance personalized medicine. However, isolating PBMCs has traditionally been a time-consuming and labor-intensive process, requiring meticulous protocols like density gradient centrifugation or red blood cell (RBC) lysis. These methods are not only tedious but can also introduce variability into experiments due to inconsistent sample handling and processing.
Whole blood undergoes dynamic changes after collection, which can lead to time-dependent changes, such as the degradation of proteins and RNA that can compromise analytical readouts if samples are not processed promptly. While chemical fixation or blood stabilization tubes can be used to preserve phenotype as well as proteomic and transcriptomic profiles, these approaches are incompatible with downstream PBMC isolation using density gradient centrifugation. This presents a challenge for workflows requiring both sample preservation and high-quality PBMC recovery.
Overcome these challenges with the EasySep™ Direct Human PBMC Isolation Kit. This cell isolation kit streamlines the isolation process, allowing researchers and laboratory professionals to rapidly and efficiently isolate PBMCs directly from blood samples in as little as 20 minutes, without the need for RBC sedimentation, centrifugation, or lysis. By minimizing hands-on time and reducing the complexity of traditional methods, the EasySep™ Direct Human PBMC Isolation Kit enables laboratory staff to focus on their analyses rather than on cumbersome sample preparation.
Learn how this unique cell isolation kit works and explore its real-world applications, from upstream sample preparation to downstream research workflows.
Why Use the EasySep™ Direct Human PBMC Isolation Kit?
- Isolate human PBMCs easily and efficiently in as little as 20 minutes without the need for lysis or centrifugation.
- Minimize handling by avoiding laborious and time-consuming methods such as density gradient centrifugation and RBC lysis.
- Obtain highly purified cells, even from older blood samples.
- Automate blood sample processing with RoboSep™ instruments to increase laboratory throughput.
- Process cells from various sources, including whole blood, cord blood, bone marrow, buffy coats, leukapheresis products, and leukocyte reduction system (LRS) cones.
How Does the EasySep™ Direct Human PBMC Isolation Kit Work?
The EasySep™ Direct Human PBMC Isolation Kit uses immunomagnetic separation technology to isolate PBMCs quickly and effectively. In this EasySep™ negative selection procedure, unwanted cells are labeled with antibody complexes and magnetic particles called EasySep™ Direct RapidSpheres™. The following unwanted cells are targeted for removal: granulocytes, platelets, and RBCs. The magnetically labeled cells are then separated from the untouched desired PBMCs by using an EasySep™ magnet and simply pouring or pipetting the desired cells into a new tube. Following magnetic cell isolation, the desired PBMCs are ready for downstream applications.
This approach eliminates the need for density gradient media, centrifugation, and RBC lysis, drastically simplifying PBMC isolation. The straightforward protocol ensures consistent results, even for laboratories with limited time or resources.
How Does EasySep™ Direct Work?
EasySep™ Direct immunomagnetically depletes red blood cells and unwanted cells in a single step without density gradient centrifugation, RBC lysis, or other pre-processing steps that can alter cellular function or increase cell isolation time. This video provides a brief introduction to this immunomagnetic platform for fast and easy cell isolation from up to 16 samples at once.
Try EasySep™ Direct in Your Own Lab!
See for yourself how EasySep™ cell separation technology provides a simpler way to isolate PBMCs with minimal handling and processing time.
See the Data: Superior PBMC Recovery, Purity, and Contaminant Removal Compared to Density Gradient Centrifugation and Column-Based Kits
The EasySep™ Direct Human PBMC Isolation Kit is designed to isolate high-quality peripheral blood mononuclear cells directly from whole blood without the need for density gradient centrifugation. In comparative studies with both density gradient centrifugation and column-based kits, this EasySep™ kit demonstrated significantly lower contamination by platelets, red blood cells, and granulocytes, along with superior mononuclear cell purity and recovery. This streamlined, column-free process minimizes debris and enhances yield, producing cleaner and more consistent PBMC preparations ideal for downstream diagnostic and research applications.
Figure 1. Isolating Cells with the 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 the EasySep™ Direct Human PBMC Isolation Kit. Cells were counted and analyzed by flow cytometry. (A) Both density gradient centrifugation and the 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 the 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 the 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 2. EasySep™ Direct Human PBMC Isolation Kit Reduces Platelet Contamination More Efficiently Than Column-Based Methods
Peripheral blood mononuclear cells (PBMCs) were isolated from 24-hour-old whole blood samples using either the EasySep™ Direct Human PBMC Isolation Kit or alternative, column-based kits. Flow cytometry analysis demonstrated that PBMCs isolated with the EasySep™ kit contained significantly fewer contaminating platelets (CD42b+), while levels of residual red blood cells (Glycophorin A+/CD45-) and granulocytes (CD66b+) were comparable between methods. Data are presented as mean ± SD (n ≥ 3 per condition).
Figure 3. EasySep™ Direct PBMC Isolation Kit Achieves Higher Recovery and Purity of Mononuclear Cells Compared to Column-Based Kits
PBMCs were isolated from 24-hour-old whole blood samples using the EasySep™ Direct Human PBMC Isolation Kit and two column-based kits designed for PBMC isolation from whole blood. Isolated cells were labeled with anti-CD45 antibodies and analyzed by flow cytometry. Cell purity was assessed by the proportion of CD45+ events among total recorded events (serving as an indicator of debris), and total recovery was quantified across isolation methods. The EasySep™ Direct PBMC Isolation Kit yielded higher purity PBMCs than both Column-Based Kits A and B. It also recovered a greater number of CD45+ mononuclear cells compared to Column-Based Kit A, with a comparable recovery to Column-Based Kit B. Data are presented as mean ± SD (n ≥ 3 per condition).
Applications Spotlight: Advancing Discovery and Diagnostics with EasySep™
Single-Cell RNA Sequencing with Fixed Blood Samples
PBMCs isolated with the EasySep™ Direct Human PBMC Isolation Kit are ideal for applications like single-cell RNA sequencing (scRNA-seq). has developed a protocol for isolating PBMCs from fixed whole blood samples using this EasySep™ kit upstream of scRNA-seq with their Chromium Fixed RNA Profiling workflow. Storage recommendations for isolated PBMCs from fixed blood are also outlined. Fixing whole blood samples allows researchers to store them for extended periods while preserving single-cell transcriptomic readouts post-draw. This flexibility is especially valuable for studies where immediate sample processing may not be feasible.
Blood Collection with Cyto-Chex® BCT Tubes
The EasySep™ Direct Human PBMC Isolation Kit was used for PBMC isolation from blood stored in Streck’s Cyto-Chex® BCT Tubes, which are blood collection tubes that maintain cellular morphology and surface antigen expression prior to analysis by flow cytometry. However, the stabilization of the whole blood samples impacts the ability to isolate PBMCs using traditional density gradient centrifugation methods. demonstrated that PBMCs could be successfully isolated from whole blood stabilized by Cyto-Chex® BCT up to 7 days post blood collection using EasySep™ Direct, while preserving cell phenotypes for downstream flow cytometric analysis. This workflow provides laboratories with the additional flexibility to transport and process samples without compromising the quality of the isolated PBMCs.
Automating PBMC Isolation with RoboSep™
Automate your PBMC isolations to minimize sample handling and free up valuable hands-on time. The EasySep™ Direct Human PBMC Isolation Kit can be used with the Dzdz™- or Dzdz™-16 instrument, automating all sample labeling and magnetic separation steps. This enables simultaneous cell isolations from up to 16 samples and increases sample throughput, laboratory output, and overall efficiency, while reducing technician time and manual handling. demonstrated that the number of PBMCs isolated from whole blood was about 2.44 times higher using RoboSep™ compared to a traditional density gradient isolation method.
Cryopreservation, Biobanking, and Thawing PBMCs
Cryopreservation:
Storage and cryopreservation of cells and tissues are important parts of the workflow for biological research. Using the appropriate cryopreservation medium, cell concentration, and freezing rate ensures that PBMCs maintain optimal viability and functionality upon thawing. With a serum-free cryopreservation medium such as cGMP CryoStor® CS10, you can ensure consistency and control according to quality standards when cryopreserving a variety of sensitive cell and sample types.
Biobanking:
Biobanking plays a crucial role in preserving high-quality PBMC samples for future research applications, including vaccine development, biomarker discovery, and drug testing. Automation has significantly enhanced biobank workflows by improving sample consistency, reducing hands-on processing time, and increasing throughput. The (NSWHSB) validated the use of the EasySep™ Direct Human PBMC Isolation Kit with the Dzdz™- platform, demonstrating its ability to efficiently isolate and cryopreserve PBMCs while maintaining high viability and recovery rates. By incorporating automation into PBMC isolation, biobanks can ensure standardized sample processing, meet regulatory requirements, and support large-scale research initiatives with minimal variability.
Thawing:
Fresh primary cells are often cryopreserved (i.e. frozen) for long-term storage in liquid nitrogen if they are not required immediately. Thawing frozen cells requires proper technique and handling to ensure optimal viability, recovery, and functionality of the cells for downstream applications. While a 37°C water bath can be used to thaw frozen vials, the ThawSTAR® CFT2 Automated Thawing System offers improved sample sterility and consistent thawing performance through a standardized thawing process.
Learn more about what makes EasySep™ Direct the easy choice for simplified cell isolation directly from whole blood.
Additional Resources for PBMC Isolation and Blood Sample Preparation
Blood Sample Preparation E-Book
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