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Miltenyi Biotec Inc.
2303 Lindbergh Street
Auburn, CA 95602
USA
Phone: +1 800 FOR MACS
+1 530 888 8871
Fax: +1 530 888 8925
E-Mail: macs@miltenyibiotec.com
Web: www.miltenyibiotec.com
For immediate technical support, use our live chat.
Connect with usOrders:
Phone: +1 866 811 4466
Fax: +1 877 591 1060
E-Mail: orderdesk@miltenyibiotec.com
Technical support - Research products:
Phone: +1 800 FOR MACS
Fax: +1 530 745 2806
E-Mail: science@miltenyibiotec.com
Technical support - Clinical products:
Phone: +1 800 FOR MACS
Fax: +1 530 745 2806
E-Mail: science@miltenyibiotec.com
Miltenyi Biotec distribution:
As a global market leader with numerous subsidiaries and distributors, Miltenyi Biotec is committed to providing our customers around the world with the highest quality products. In addition to direct selling in more than 20 countries in North America, Europe and Asia/Pacific, Miltenyi Biotec also provides support for our customers through an extensive distributor network covering dozens of additional countries.
Our local employees are always happy to answer your questions. Highly trained and experienced teams in your country can provide quick, helpful, and comprehensive support.
Miltenyi Biotec Inc.
2303 Lindbergh Street
Auburn, CA 95602
USA
Phone: +1 800 FOR MACS
+1 530 888 8871
Fax: +1 530 888 8925
E-Mail: macs@miltenyibiotec.com
Web: www.miltenyibiotec.com
For immediate technical support, use our live chat. Connect with us
Orders
Phone: +1 866 811 4466
Fax: +1 877 591 1060
E-Mail: orderdesk@miltenyibiotec.com
Technical support - Research products
Phone: +1 800 FOR MACS
Fax: +1 530 745 2806
E-Mail: science@miltenyibiotec.com
Technical support - Clinical products
Phone: +1 800 FOR MACS
Fax: +1 530 745 2806
E-Mail: science@miltenyibiotec.com
As a global market leader with numerous subsidiaries and distributors, Miltenyi Biotec is committed to providing our customers around the world with the highest quality products. In addition to direct selling in more than 20 countries in North America, Europe and Asia/Pacific, Miltenyi Biotec also provides support for our customers through an extensive distributor network covering dozens of additional countries.
Granulocytes are myeloid cells that play key roles in innate immune responses. They are characterized by the presence of cytoplasmic granules and a poly-lobed nucleus, which gives them the name polymorphonuclear leukocytes (PMN, PML or PMNL).
Generated and differentiated in bone marrow, granulocytes have a limited life span once in the periphery and thus, cannot be maintained in culture for prolonged periods of time. They play a well-known role in inflammation and allergy, but are also capable of antigen presentation and immune response modulation (PMID: 23007469). Among the first responders at site of inflammation, granulocytes are equipped with preformed granules filled with an array of potent effector molecules to fight invading pathogens. Interestingly, depending on their activation status and environment, granulocytes have also been implicated in disease. For example, neutrophils can promote tumor growth, modulate their extracellular matrix or enhance tumor angiogenesis (PMID: 26819959, 23485549, 28099862, 27558343).Dendritic cells (mouse)
Granulocyte research (brochure)
At a glance: granulocyte subtypes
Cell subtype | Frequency | Markers | Function |
Neutrophils | Frequencies of granulocytes are variable and depend on tissue and mouse strain | CD11b, Ly-6G, Gr-1, CD115- |
|
Eosinophils | CD11b, Ly-6C, Siglec-F, CD193 (CCR3) |
| |
Basophils | CD11b, CD49b, FcεRIα, CD200R3; CD117- |
|
Miltenyi Biotec has developed numerous products for the magnetic separation of granulocytes. Both positive selection and depletion strategies can be used to isolate granulocytes, depletion is preferred when working with bone marrow, and positive selection when working with other tissues.
For details on MACS® Cell Separation Technology, see the MACS Handbook chapter Magnetic Cell Separation .Magnetic cell separation
At a glance: Kits and reagents for the separation of neutrophils
Cell subtype | Starting material | Isolation strategy | Comments | Automation | Product |
---|---|---|---|---|---|
Neutrophils | Lymphoid tissues | Positive selection of target cells | Yes* | Anti-Ly-6G MicroBead Kit, mouse | |
Neutrophils | Bone marrow or blood cell suspensions | Depletion of non-target cells | Untouched neutrophils | Yes* | Neutrophil Isolation Kit, mouse |
Neutrophils and eosinophils | Blood and other tissues | Positive selection of target cells | Yes* | Siglec-F MicroBeads, mouse | |
*Automation options range from fully automated benchtop solutions such as the autoMACS® Pro Separator to high-throughput platforms such as the MultiMACS™ Cell24 Separator Plus or MultiMACS X. |
In contrast to human neutrophils which are easily harvested in large numbers from blood (PMID: 18432834), the small volume of mouse blood makes isolating sufficient numbers of neutrophils to perform downstream analyses rather difficult. Bone marrow is a convenient alternative source of neutrophils and their precursors, both at steady-state under homeostatic conditions and during activation under a variety of infectious and non-infectious inflammatory conditions. Bone marrow neutrophils are functionally similar to blood neutrophils in mice, are reported to survive longer in culture (PMID: 14694182), and can become activated. The number of cells that can be obtained is compatible with the majority of immunological assays and adoptive transfer studies, making bone marrow a useful reservoir for isolating neutrophils in a variety of mouse strains and disease models.
The Neutrophil Isolation Kit, mouse enables isolation of untouched neutrophils from bone marrow. Non-target cells are labeled with a cocktail of biotinylated antibodies for subsequent depletion, resulting in a highly pure population of isolated neutrophils, which are viable and ready for functional studies (PMID: 28783679). This kit can also be used to isolate neutrophils from blood cell suspensions.Unseparated fraction
Isolated neutrophils
Murine neutrophils isolated from bone marrow. Neutrophils were isolated from C57BL/6 bone marrow cells using the Neutrophil Isolation Kit, mouse, an LS Column, and a MidiMACS™ Separator. Cells were fluorescently stained with Anti-Ly-6G-PE and CD11b-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
MACS Cell Separation - Select the best (brochure)
Granulocyte research (brochure)
Cap'n T Cell explains what makes MACS® Technology unique and superior and why it is the best choice for isolating your cells.
At a glance: Kits and reagents for the separation of eosinophils
Cell subtype | Starting material | Isolation strategy | Comments | Automation | Product |
---|---|---|---|---|---|
Neutrophils and eosinophils | Blood and other tissues | Positive selection of target cells | Yes* | Anti-Siglec-F MicroBeads, mouse | |
*Automation options range from fully automated benchtop solutions such as the autoMACS® Pro Separator to high-throughput platforms such as the MultiMACS™ Cell24 Separator Plus or MultiMACS X. |
Blood
Before separation
Siglec-F+ cells
Lung
Before separation
Siglec-F+ cells
Eosinophils isolated from mouse blood and mouse lung tissue. Anti-Siglec-F MicroBeads, two MS Columns, and a MiniMACS Separator were used to isolate eosinophils from mouse blood and mouse lung. Isolation from blood resulted in purely enriched eosinophils, which were stained with Anti-Siglec-F-APC and Gr-1 VioGreen for flow cytometry analysis. Isolation from lung resulted in a mixture of eosinophils and Siglec-F+ alveolar macrophages. These cells were stained with Anti-Siglec-F-PE and CD11c-PE-Vio770 for flow cytometry analysis. Analyses were performed on the MACSQuant® Analyzer, and cell debris and dead cells were excluded based on scatter signals and propidium iodide fluorescence. Equal event numbers are shown for the respective dot plots.
At a glance: Kits and reagents for culturing bone marrow to generate eosinophils
Use | Comments | Product |
---|---|---|
Supplement | Expansion of progenitor cells | Mouse SCF |
Supplement | Mouse Flt3-Ligand | |
Supplement | Mouse IL-5 |
Cell culture
At a glance: Markers for the discrimination of granulocyte subtypes (PMID: 26793192)
Surface or intracellular markers | |||||||||
CD11b | CD49b | FcεRIα | CD177 | Ly-6G | Ly-6C | CD115 | Siglec-F | CD200R3 | |
Neutrophils | + | – | – | – | + | Lo | – | – | – |
Eosinophils | + | – | – | – | – | Lo | – | + | – |
Basophils | Population smear gradually going from dim to positive | + | + | – | – | – | – | – | + |
REAfinity Recombinant Antibodies (brochure)
Granulocyte research (brochure)