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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.
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Tumor-infiltrating leukocytes research (brochure)
Optimal storage of murine tumors in MACS® Tissue Storage Solution (application note)
The amount and composition of TILs is highly variable, complicating the analysis of individual subpopulations. When working with large cohort sizes, even immunophenotyping of TILs by flow cytometry is time consuming and data processing highly work intensive. Therefore, pre-enrichment of TILs is desirable to increase the sensitivity of analysis and save time and effort during downstream analysis such as sequencing and flow cytometry. To this end, Miltenyi Biotec offers optimized MicroBeads for tumor tissue samples.
Isolation of CD4+/CD8+ TIL subpopulation with REAlease Technology followed by dead cell removal improves cell vitality
Using the REAlease CD4/CD8 (TIL) MicroBead Kit, human directly on dissociated tumor tissue provides highly pure CD4+/CD8+ human TILs and delivers MicroBead- and label-free cells. Isolated cells are suitable for further processing, e.g., with the MicroBead-based Dead Cell Removal Kit, human, thereby improving the vitality of CD4+/CD8+ TILs from 80% to nearly 90%.
Magnetic cell separation
One major goal in immunotherapies is to understand how the composition of tumor-infiltrating myeloid and lymphoid cells contributes to patient stratification. However, TIL numbers can be very low and small subpopulations might be lost in background noise, particularly in single-cell analyses. Therefore, serial sorting of TIL subpopulations is highly advantageous in order to investigate the immune cell composition.
Background-free flow analysis of TILs
Using hybridoma-derived antibodies to identify TILs leads to a gross overestimation of the frequency of immune cell subpopulations present in the tumor. This experimental artifact is most likely caused by unspecific binding of hybridoma-derived antibodies to FcγRs on immune cells, as it is significantly reduced when using an FcR blocking reagent.
In contrast, assessment of TIL frequency using REAfinity™ Recombinant Antibodies is unchanged in the presence or absence of FcR blocking, providing a more exact analysis of respective cell populations, even in the absence of FcR blocking.
REAfinity Antibodies are highly specific recombinant antibodies that provide superior lot-to-lot consistency and purity compared to mouse or rat hybridoma-derived monoclonal antibodies.
Phenotypic characterization of TILs by flow cytometry can be performed using optimized panels of fluorochromes-conjugated REAfinity Recombinant Antibodies.
Panel 1 | Clone | Fluorochrome | Panel 2 | Clone | Fluorochrome |
---|---|---|---|---|---|
CD8β | REA793 | VioBlue® | CD8β | REA793 | VioBlue |
Viobility | 405/520 | Viobility | 405/520 | ||
Lag-3 | REA776 | VioBright™ 515 | CD103 | REA789 | VioBright 515 |
TIM-3 | REA602 | PE | CXCR3 | REA724 | PE |
CD4 | REA604 | PE-Vio® 615 | CD4 | REA604 | PE-Vio 615 |
PD-1 | REA802 | APC | CD39 | REA870 | APC |
CD44 | REA664 | APC-Vio 770 | CD44 | REA664 | APC-Vio 770 |
Background-free analysis of mouse TILs (application note)
Discover REAfinity Antibodies
The tumor microenvironment (TME) is inherently complex due to the highly variable cell composition as well as to interactions of thousands of proteins involved in metastasis formation. Thus, an in-depth analysis of tumor tissues requires the examination of a plethora of parameters in order to decipher the underlying principles. Currently available techniques can provide only a very limited perspective on TME complexity.
Miltenyi Biotec’s MICS (MACSimaTM Imaging Cyclic Staining) technology impressively overcomes these limits as it allows the fluorescence microscopic analysis of hundreds of markers on a single sample, without any harm, in a fully automated manner.
MACSima Imaging Platform
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