Magnetic activated cell sorting (macs)

Manufactured by Miltenyi Biotec

Sourced in Germany, United States, United Kingdom, Canada, Japan, Italy, Singapore

MACS is a magnetic cell separation technology developed by Miltenyi Biotec. It enables the efficient isolation and enrichment of target cells from complex biological samples. The core function of MACS is to provide a gentle, reliable, and scalable method for cell separation and purification.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Magnetic cell sorting (66 citations) Magnetic sorting (25 citations) MACS sorting (18 citations)

838 protocols using magnetic activated cell sorting (macs)

Magnetic Enrichment of Biotinylated Protein Targets

Check if the same lab product or an alternative is used in the 5 most similar protocols

K562 cells stably expressing the sunCAS9 system carrying the pooled sgRNA CSR libraries (TM1; TM2+) were expanded and 50 million cells per screen and library were harvested, washed three times with cold MACS (Miltenyi) buffer, and resuspended in 2 ml MACS buffer in a sterile 5 ml Eppendorf tube. Cells were then labeled with magnetic streptavidin microbeads complexed with biotinylated bait protein (50 µl streptavidin microbeads; 1 µM biotinylated protein), mixed and incubated at 4°C for 30 min (tumbling). After labeling, cells were washed twice with cold MACS buffer (300× g, 10 min), resuspended in 1 ml MACS buffer and passed through a 40 µM cell strainer, to obtain a single-cell suspension, directly onto the LS-Column (Miltenyi) for magnetic bead separation according to the manufacturer’s protocol. Briefly, after applying the labeled cells onto the LS-Column, unlabeled cells pass through and are discarded while labeled cells are retained in the magnetic field, the LS-Column is washed three times with 3 ml of ice-cold MACS buffer. For elution of positively selected cells, the column is removed from the separator (magnet) and the magnetically labeled cells are flushed into a 15 ml Falcon tube with fresh media (RPMI complete), washed once, resuspended, and transferred to a T25 culture flask for expansion.

Siepe D.H., Henneberg L.T., Wilson S.C., Hess G.T., Bassik M.C., Zinn K, & Garcia K.C. (2022). Identification of orphan ligand-receptor relationships using a cell-based CRISPRa enrichment screening platform. eLife, 11, e81398.

+ Open protocol

+ Expand

hiPSCs-MSCs Phenotypic Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols

The hiPSCs-MSCs at passage 3 were harvested. One million cells were suspended in 100 µl buffer that consisted of 0.5% bovine serum albumin (BSA; Sigma-Aldrich; Merck Millipore) and 2 mM EDTA (Sunshine Biotechnology Co., Ltd., Nanjing, China). Subsequently, 10 µl 1:10 diluted fluorescein isothiocyanate (FITC)-coupled antibodies recognizing CD11b (130-098-778), CD105 (130-098-778), CD90 (130-097-930), CD45 (130-098-043) and CD34 (130-098-142) (MACS; Miltenyi Biotec, Bergisch Gladbach, Germany) were added. In addition, 1:10 diluted mouse IgG1 (130-104-562) and mouse IgG2a (no. 130-098-877) antibodies (MACS; Miltenyi Biotec) were used as isotype controls. Incubation for 10 min incubation in the dark at 4°C was performed. The cells were then washed with buffer containing phosphate-buffered saline, pH 7.2, 0.5% BSA, and 2 mM EDTA by diluting MACS BSA Stock Solution (130-091-376) 1:20 with autoMACS Rinsing Solution (130-091-222) (MACS; Miltenyi Biotec). Then, the cells were centrifuged at 300 × g for 10 min at 4°C and resuspended in 500 µl of the aforementioned buffer for analysis by flow cytometry (BD FACSCalibur, BD Biosciences, Franklin Lakes, NJ, USA). The data was analyzed using Flowjo 7.6 sofrware (BD Biosciences).

Xu X., Shi D., Liu Y., Yao Y., Dai J., Xu Z., Chen D., Teng H, & Jiang Q. (2017). In vivo repair of full-thickness cartilage defect with human iPSC-derived mesenchymal progenitor cells in a rabbit model. Experimental and Therapeutic Medicine, 14(1), 239-245.

+ Open protocol

+ Expand

Establishment of Novel ENKTL Cell Line

Check if the same lab product or an alternative is used in the 5 most similar protocols

The novel ENKTL cell line LY was established from PDX-#1 murine model. After two PDX passages, separation of CD56⁺ cells were performed by positive selection cell sorting methods utilizing anti-human CD56 antibody-based magnetic beads (MACS, Miltenyi Biotec) from splenic mononuclear cells of the PDX-#1 murine model (anti-human CD56⁺ population: 73.4% before MACS isolation versus 94.58% after MACS isolation). The purified CD56⁺ cells were subjected to cell culturing under the same culture conditions as SNK-6 cells in the supply of human recombinant IL-2 until the cells constantly doubled their number within 24 h. The detailed information is shown in Figure S1. The morphology of LY cell cells is similar to that of SNK-6 cells, exerting several cell clumps in suspension and medium to large cell size with large nuclei and abundant cytoplasm. Cell-surface marker analysis by flow cytometry revealed that LY cells are HLA-DR⁺CD3⁻CD16⁻CD56⁺, indicating an NK phenotype. Finally, the STR patterns of the LY cells did not match those of any cell lines in the public cell banks, such as the ATCC, DSMZ, or JRCB, and did not contaminate with murine cells. Hence it is concluded that LY is a new ENKTL cell line.

Liu C., Ding X., Li G., Zhang Y., Shao Y., Liu L., Zhang W., Ma Y., Guan W., Wang L., Xu Z., Chang Y., Zhang Y., Jiang B., Yin Q, & Tao R. (2023). Targeting Bcl-xL is a potential therapeutic strategy for extranodal NK/T cell lymphoma. iScience, 26(8), 107369.

+ Open protocol

+ Expand

Isolation and Sorting of Human Regulatory T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

CD4⁺ T cells were separated from PBMC from healthy controls using pan T‐cell isolation kit and magnetic‐activated cell sorting (Miltenyi Biotec, Bergisch Gladbach, North Rhine‐Westphalia, Germany). Enriched T cells were subsequently stained with CD25‐PE mAb and CD25⁺ T cells positively selected using anti‐PE magnetic beads and MACS (Miltenyi Biotec, Bergisch Gladbach, North Rhine‐Westphalia, Germany). To obtain high purity Treg, CD25^highCD127⁻ T cells were subsequently sorted in FACSAria III (BD Biosciences, Franklin Lakes, New Jersey, USA). The purity of the sorted Treg subsets was assessed by staining the cells with FoxP3‐APC mAb. CD49f^‐ and CD49f^high cells were further separated from the total fraction of Treg by FACS‐based sorting based on CD49f FMO controls. Over 95% of purity was detected in all cases analysed. The remaining CD25⁻ fraction was assessed as conv CD4⁺ populations.

Weerakoon H., Straube J., Lineburg K., Cooper L., Lane S., Smith C., Alabbas S., Begun J., Miles J.J., Hill M.M, & Lepletier A. (2021). Expression of CD49f defines subsets of human regulatory T cells with divergent transcriptional landscape and function that correlate with ulcerative colitis disease activity. Clinical & Translational Immunology, 10(9), e1334.

+ Open protocol

+ Expand

Cardiac Stem Cell Isolation and Differentiation

Check if the same lab product or an alternative is used in the 5 most similar protocols

CSCs were isolated from adult transgenic and wt hearts by enzymatic methods.^{35 (link), 40 (link)} For CD45⁻c-Kit⁺ cell purification, myocyte-depleted small cardiac cells were incubated with microbeads conjugated with anti-mouse CD45 antibody (Miltenyi Biotec S.r.l., Calderara di Reno (BO), Italy) and removed from the preparation by magnetic-activated cell sorting (Miltenyi). From the CD45⁻ fraction, the c-Kit⁺ (CD45⁻) cardiac cells were enriched through incubation with a microbeads-conjugated mouse monoclonal antibody against c-Kit (Miltenyi) and separated by magnetic-activated cell sorting.
Freshly isolated CD45⁻c-Kit⁺ cardiac cells were cultured on gelatin-coated dishes in CSC growth medium^{35 (link), 40 (link)} before clonogenic, spherogenic and BrdU assays.
Cardiosphere myogenic differentiation was performed as previously described.^{35 (link)}Endothelial differentiation was obtained by culturing the CSC for 3–10 days in MEM Alpha (Life Technologies), 10% ESQ-FBS (Life Technologies), 1 μM dexamethasone, 50 μM/ml ascorbic acid, 10 mM β-glycerophosphate (all from Sigma-Aldrich, Milano, Italy) and 10 ng/ml VEGF (PeproTech EC Ltd., London, UK). LY294002 (10 μM, Calbiochem, San Diego, CA, USA) and UO126 (10 μM, Cell Signalling Technology, Danvers, MA, USA) were added to the culture for AKT and ERK1/2 inhibition.

Di Siena S., Gimmelli R., Nori S.L., Barbagallo F., Campolo F., Dolci S., Rossi P., Venneri M.A., Giannetta E., Gianfrilli D., Feigenbaum L., Lenzi A., Naro F., Cianflone E., Mancuso T., Torella D., Isidori A.M, & Pellegrini M. (2016). Activated c-Kit receptor in the heart promotes cardiac repair and regeneration after injury. Cell Death & Disease, 7(7), e2317-.

+ Open protocol

+ Expand

Isolation and Purification of Regulatory T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

CD4 + T cells were separated from healthy controls PBMC using pan T cell isolation kit and magnetic-activated cell sorting (Miltenyi Biotec, USA). Enriched T cells were subsequently stained with CD25-PE mAb and CD25 + T cells positively selected using anti-PE magnetic beads and MACS (Miltenyi Biotec, USA). To obtain high purity Treg, CD25 high T cells were stained with CD3-APCe780, CD4-BV711 and CD127-BV786 mAbs (BD Biosciences), and zombie aqua live/dead marker (Biolegend, USA) and sorted in FACSAria III (BD Biosciences, USA). The purity of the sorted Treg subsets was assessed by staining the cells with FoxP3-APC mAb using the FoxP3 Fix/Perm Buffer Set for nuclear staining (Biolegend, USA). Over 95% of purity was detected in all cases analyzed.
The CD25 -fraction was assessed as conv CD4 + populations.

Weerakoon H., Straube J., Lineburg K.E., Cooper L., Lane S., Smith C., Alabbas S., Begun J., Miles J.J., Hill M.M., & Lepletier A. (2021). Expression of CD49f defines subsets of human regulatory T cells with divergent transcriptional landscape and function that correlate with ulcerative colitis disease activity.

+ Open protocol

+ Expand

Isolation of Tumor Myeloid Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lung tumour biopsies or healthy tissues were minced in RPMI medium containing 0.1% collagenase type I, 0.2% dispase type I and DNase I 100 U/ml (60 minutes at 37°C), passed through a 19 G needle and passed through a 70 and 40 μm cell strainer. After RBC lysis, cells were resuspended in FACS buffer (PBS containing 2% FBS and 2 mM EDTA) and counted. Myeloid population, enriched using CD11b-conjugated magnetic beads (MACS, Miltenyi Biotec) and separated through magnetic column (MACS, Miltenyi Biotec), was stained with anti-CD66b (G10F5, BD Pharmingen) for 20 minutes on ice and sorted with FACS Aria I (BD Bioscience). Cells were counted and resuspended in RLT buffer (Qiagen) for RNA extraction.

Finisguerra V., Di Conza G., Di Matteo M., Serneels J., Costa S., Thompson A.A., Wauters E., Walmsley S., Prenen H., Granot Z., Casazza A, & Mazzone M. (2015). MET is required for the recruitment of anti-tumoural neutrophils. Nature, 522(7556), 349-353.

+ Open protocol

+ Expand

Isolation of Tumor Myeloid Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

NK Cell Enrichment and Labeling

Check if the same lab product or an alternative is used in the 5 most similar protocols

For in vitro experiments, activated NK cells were enriched from HD PBMCs by CD56-positive immunomagnetic separation (MACS, Miltenyi). NK cells were then fluorescently labelled with 3 µM of eFluor670 (eBioscience), following the manufacturer’s instructions.
For in vivo experiments, NK cells from freshly isolated PBMCs of HDs were enriched by CD3-negative immunomagnetic separation (MACS, Miltenyi) before NK cell expansion.

Lanuza P.M., Alonso M.H., Hidalgo S., Uranga-Murillo I., García-Mulero S., Arnau R., Santos C., Sanjuan X., Santiago L., Comas L., Redrado S., Pazo-Cid R., Agustin-Ferrández M.J., Jaime-Sánchez P., Pesini C., Gálvez E.M., Ramírez-Labrada A., Arias M., Sanz-Pamplona R, & Pardo J. (2022). Adoptive NK Cell Transfer as a Treatment in Colorectal Cancer Patients: Analyses of Tumour Cell Determinants Correlating With Efficacy In Vitro and In Vivo. Frontiers in Immunology, 13, 890836.

+ Open protocol

+ Expand

Isolation of Leukemic and Healthy Hematopoietic Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bone marrow (BM) and blood samples were collected at the time of diagnosis, CR and relapse from AML patients after informed consent in accordance with the declaration of Helsinki and the local ethics committee of Saint-Antoine hospital. Mononuclear cells were obtained after ficoll separation and cryopreserved at the tumour bank of Saint-Antoine Hospital, Paris. Fresh or frozen samples were used for further cellular and molecular experiments. BM cells were sorted to enrich the samples in leukaemic or non-leukaemic progenitor cells. CD34⁺, CD38⁺, CD34⁻ or CD34⁺CD38^- cell sorting strategies were established according to initial leukaemia immuno-phenotype. Blood cells from CR were sorted to isolate the CD3⁺ fraction. Immuno-magnetic cell sorting was performed using CD3, CD34 or CD38 MicroBeads and columns (MACS, Miltenyi Biotec) according to the manufacturer's protocols.
Umbilical cord blood samples were collected from healthy new-borns with mothers' consent in accordance with the institutional review boards of the Etablissement Français du Sang, Créteil France, and the Institut National de la Santé et de la Recherche Médicale, Paris, France. CD34⁺ cells were isolated using a dextran/ficoll based procedure followed by immuno-magnetic separation (MACS, Miltenyi Biotec).

Hirsch P., Zhang Y., Tang R., Joulin V., Boutroux H., Pronier E., Moatti H., Flandrin P., Marzac C., Bories D., Fava F., Mokrani H., Betems A., Lorre F., Favier R., Féger F., Mohty M., Douay L., Legrand O., Bilhou-Nabera C., Louache F, & Delhommeau F. (2016). Genetic hierarchy and temporal variegation in the clonal history of acute myeloid leukaemia. Nature Communications, 7, 12475.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!