Discover this step-by-step workflow for isolation and characterization of human as well as mouse B cells and their subsets. MACS® Technology allows for fast and easy B cell isolation and optimized flow cytometry reagents enable reliable B cell analysis.
B cells in SARS-CoV-2
Step-by-step protocols and application note for analysis, enrichment, and expansion of SARS-CoV-2-specific B cells
We have a diverse range of solutions for B cell separation. These solutions include untouched isolation, positive isolation, isolation directly from different blood products and REAlease® Technology, to offer you a streamlined method for fast and efficient B cell isolation. Whatever your subset of interest or starting material, we have the right tool for you.
Magnetic B cell separation with MACS® Technology
MACS Technology offers a solution to obtain a highly purified population separated by positive selection or untouched cells from whole blood, untouched B cells and B cell subsets from human PBMCs.
Label-free B cell isolation
REAlease Technology allows for magnetic cell isolation with subsequent removal of any beads and labels from your B cells of interest. Clever utilization of recombinant antibody fragments ensures cell surface labeling in just a few steps.
The technology relies on recombinantly engineered antibody fragments for cell surface labeling, ensuring highly reproducible results. The REAlease Biotin Complex binds to the target cells. Labeling of the REAlease Biotin Complex with Anti-Biotin MicroBeads promotes magnetic isolation of these cells. Following cell separation, both MicroBeads and the REAlease Biotin Complex are gently removed, leaving the cells bead- and label-free. Enriched cells are then suitable for magnetic relabeling and any application where label-free cells are essential.
REAlease MicroBeads
- Bead-free cells suited for a second positive selection
- Label-free cells for any downstream application
- Recombinant antibody fragments for reproducible results
Comparison of activation marker expression of label free B cells using REAlease MicroBeads and untouched isolation
- MACS MicroBeads – MicroBeads are available with many human, mouse, rat and non-human primate specificities
- B Cell Isolation Kits – untouched isolation of human and mouse B cells and B cell subsets
Whatever your isolation method of choice, with MACS Technology you obtain non-activated
B cells compatible for downstream applications.
Untouched human B cell isolation
Our B Cell Isolation Kits enable you to isolate highly pure, untouched B cells and B cell subsets from human PBMCs.
Find the best kit suited to your research:
| Isolated cells | Isolated procedure | Isolation Kit |
|---|---|---|
| Activated B cells, malignant B cells (if there are any) and plasma cells | Depletion of unwanted cells | Pan B Cell Isolation Kit, human |
| Resting, immature and naive B cells, CD43+ cells depleted | Depletion of unwanted cells | B Cell Isolation Kit II, human |
| CD27+ memory B cells | Depletion of unwanted cells and subsequent positive selection of target cells | Memory B Cell Isolation Kit, human |
| IgM expressing memory B cells | Depletion of unwanted cells and subsequent positive selection of target cells | IgM+ Memory B Cell Isolation Kit, human |
| IgG expressing memory B cells | Depletion of unwanted cells and subsequent positive selection of target cells | IgG+ Memory B Cell Isolation Kit, human |
| Switched Memory B cells | Depletion of unwanted cells | Switched Memory B Cell Isolation Kit, human |
| Naive B cells *Please note that double negative memory B cell will not be depleted with this Kit | Depletion of unwanted cells | Naive B Cell Isolation Kit II, human |
| B cells from B-CLL or other B cell malignancies CD43+ cells are not depleted | Depletion of unwanted cells | B-CLL Cell Isolation Kit, human |
B cell isolation directly from blood products
StraightFrom Technology ensures isolated B cells directly from blood products whilst saving time. The StraightFrom® MicroBeads provide magnetic cell separation directly from human blood products, including, whole blood, buffy coat, leukoreduction system chambers (LRSCs) and leukopaks. CD19+ and CD138+ B cells can be purified using the autoMACS® Pro Separator, the MultiMACS™ Cell24 Separator Plus, or manual separators.
Isolation of human CD19 positive cells that were isolated from whole blood using the StraightFrom® LRSC CD19 MicroBeads Kit.
Cells were fluorescently stained with CD20‑APC, as well as CD45‑VioBlue®, and analyzed by flow cytometry using the MACSQuant® Analyzer.
Scientific Poster
Isolation of functional cells straight from buffy coat, LRSC, and Leukopak®
Maike Schlemminger, Silke Weber-Lohmann, Martin Kolnik, Zhong Yu, Susanne Höher-Peters, and Gregor Winkels
Untouched B cell isolation from blood products using MACSxpress® Technology
For smaller starting material volumes the MACSxpress Technology isolates viable and functional B cells quickly and conveniently from blood and blood products. This cell isolation technology is based on micro-sized MACSxpress Beads, which allow the removal of non-target cells by immunomagnetic depletion using the MACSxpress Separator. All without the requirement of density gradient centrifugation or red blood cell (RBC) lysis.
MACSxpress B Cell Isolation Kits
- Untouched cell isolation from whole blood, buffy coat, or LRSC in 25 minutes
- No density gradient centrifugation or RBC lysis needed
- Flexible sample volumes from 2–30 mL
- Easy and fast protocols with only a few handling steps
- No cell activation and no effects on cell biology
Microchip-based sorting of B cells
For the isolation of rare B cell sub-populations the The MACSQuant® Tyto® Cell Sorter is revolutionizing cell sorting. Our patented microchip-based technology opens up new possibilities with high-speed multiparameter cell sorting in the safety of a fully enclosed and sterile cartridge system, the MACSQuant Tyto Cartridge. This enables gentle sorting of B cells, leading to high target cell purity and viability of theses sensitive cells.
MACSQuant® Tyto®
- Gentle cell sorting in a closed cartridge system without sheath fluid
- Engineered for operator safety with no aerosols or droplet formation
- Intuitive handling suitable for any lab professional
- High purity and viability of rare subsets
Having trouble culturing B cells?
Due to the low frequency of B cells in peripheral blood, ex vivo cultivation is an ideal option to increase B cell numbers; either to study their biology, or transform them and analyze their antibody production as well as specificity. Often, this can prove difficult, and many of the challenges associated with this can be overcome using our B Cell Expansion Kit, human. The formulation offers the perfect solution to expand and activate human B cells from various sources. This enables expansion of B cells that are fully functional and ready for any downstream application.
B cell expansion
Reliable activation and expansion of B cells is a clear requirement for effective downstream experiments. The B Cell Expansion Kit, human is optimized for primary B cell expansion in vitro. The B Cell Expansion Kit consist of a recombinant CD40L multimer, a cross-linking antibody, and IL-4. Cross-linking of CD40 expressed on B cells leads to cell activation and proliferation. Combining CD40L together with IL-4, which is a known growth factor for activated B cells, has been shown to be crucial for effective activation and proliferation of B cells.
Expansion rate
- Up to 10-fold expansion of B cells in vitro within 14 days
- Standardized and reproducible system
Frequency of B cell subsets after 14 day culture
Expanded B cells display an activated phenotype with a distribution of memory and plasma cells similar to the original fraction. The amount of naive B cells decreased under all culture conditions on day 14. This is in line with reports describing that naive B cells have a shorter survival rate.
Phenotype of expanded B cells using the B Cell Expansion Kit, human
- Similar distribution of plasma cells, naive, and memory B cells (switched and non-switched)
- At day 14, all showed a reduction of the naive B cell subset
- The amount of memory cells was similar in the expanded cell population
Scientific poster
A standardized and reproducible method for expansion and activation of human primary B cells
Aparajita Singh, Susanne Bethke, Daniela Vorholt, and Andrzej Dzionek
As the final step in many workflows, flow analysis plays an important part in various B cell applications. The following flow reagents can help you make sure that the data you generate on your flow cytometer is reliable and consistent:
- REAfinity™ Recombinant Antibodies – recombinantly generated antibodies that ensure lot-to-lot consistency and eliminate background signal
- MACSplex™ Cytokine Kits – analysis of multiple soluble cytokines in one sample using any standard flow cytometer
- Cytokine Secretion Assay – Detection Kits – flow analysis of cytokine secretion at a detection level of one in a million
- Build your own B cell flow panel with our Flow Panel Builder
Scientific poster
Increased sensitivity of flow cytometry for detection of minimal residual disease and circulating tumor cells in B Non-Hodgkin Lymphoma
Christiane Siewert, Karin Hartmann, and Thomas Nebe
MBTP 27: Immunophenotyping of B cells from human buffy coat using flow cytometry
- Keywords
- Flow cytometry and cell analysis
- Immunology
- human
- B cells
- buffy coat
- Antibody panel
- Application protocol
Expansion of isolated B cells with the B Cell Expansion Kit, human
- Keywords
- Cell culture
- Immunology
- lymphocytes
- B cells
- human
- Application protocol
Enrichment of SARS-CoV-2–specific B cells using recombinant SARS-CoV-2 proteins
- Keywords
- Cell separation
- Immunology
- lymphocytes
- B cells
- human
- MicroBeads
- Application protocol
Multicolor flow cytometric analysis of B cell subsets from mouse spleen
- Keywords
- Sample preparation
- Flow cytometry and cell analysis
- Immunology
- mouse
- spleen
- B cells
- Application protocol
Analysis of SARS-CoV-2–specific B cells using recombinant SARS-CoV-2 proteins
- Keywords
- Flow cytometry and cell analysis
- Immunology
- lymphocytes
- B cells
- human
- Antibodies
- MicroBeads
- Application protocol
Microchip-based cell sorting and expansion of SARS-CoV-2–specific B cells using the MACSQuant® Tyto® Cell Sorter and the B Cell Expansion Kit
- Keywords
- Immunology
- Flow cytometry and cell analysis
- human
- B cells
- Application protocol
