CD4+CD25− naïve T cells, B cells, and CD8+ T cells were isolated from blood samples and/or CxCa tumour tissues using MACS kits (Miltenyi nos. 130-094-131, 130-091-151 and 130-096-495 respectively) as per the manufacturer’s instructions. Naïve T cells were additionally flow-sorted to obtain a highly pure CD4+CD25− population.
Macs kits
Manufactured by Miltenyi Biotec
Sourced in Germany
MACS kits are a series of magnetic cell separation products developed by Miltenyi Biotec. They are designed to isolate and enrich specific cell populations from complex biological samples, such as blood, tissue, or cell cultures. The core function of MACS kits is to enable efficient and reliable cell separation through the use of magnetic microbeads conjugated with antibodies that bind to target cell surface markers.
Automatically generated - may contain errors
Lab products found in correlation
8 protocols using macs kits
1
Isolation of Immune Cell Subsets
2
Isolation and Culture of Human PBLs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human primary peripheral blood lymphocyte (PBL) T-cells and other immune cell subtypes were isolated from buffy coats obtained from the blood transfusion services at National University Hospital and Health Sciences Authority, Singapore using Lymphoprep™ (Axix Shield) density gradient centrifugation or using MACS kits (Miltenyi Biotec). Experiments were approved by Nanyang Technological University Institutional Review Board (IRB-2014-09-007). HuT78 T-cell line was obtained from the American Type Culture Collection. Cells were cultured in Gibco™ RPMI1640 medium supplemented with 2 mM l -glutamine, 1 mM sodium pyruvate, 10% fetal calf serum and antibiotics (penicillin and streptomycin) as described previously (17 (link), 18 (link)).
3
Isolation and Expansion of Primary T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
C57BL/6J and BALB/c mice were used as primary mouse T cell donors for polyclonal T cell expansion studies and C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-I) mice were used for antigen-specific T cell expansion studies. T cells were obtained from the spleen and isolated using pan T cell or CD8a+ T cell isolation MACS kits (Miltenyi Biotec) to obtain CD3+ T cells or CD8+ T cells for polyclonal and antigen-specific studies T cell expansion studies, respectively.
Primary human T cells were obtained from de-identified leukoreduction collars (Brigham and Women’s Hospital Specimen Bank) and used within 24 hours of initial collection (stored at RT). PBMCs were enriched from leukoreductions in a Ficoll gradient, then isolated using pan T cell or CD8+ T cell isolation MACS kits to obtain CD3+ or CD8+ T cells for polyclonal or antigen-specific studies, respectively.
Primary human T cells were obtained from de-identified leukoreduction collars (Brigham and Women’s Hospital Specimen Bank) and used within 24 hours of initial collection (stored at RT). PBMCs were enriched from leukoreductions in a Ficoll gradient, then isolated using pan T cell or CD8+ T cell isolation MACS kits to obtain CD3+ or CD8+ T cells for polyclonal or antigen-specific studies, respectively.
4
Isolation and Expansion of Primary T Cells
5
Splenocyte and CD4+ T cell isolation and activation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Splenocytes were directly obtained from the spleens after removing the red cells by BD Pharm lyse™. CD4+ T cells were purified from the spleens using MACS kits (Cat: 130-049-201, Miltenyi Biotec, Bergisch Gladbach, Germany). The splenocytes and CD4+ T cells were cultured in RPMI 1640 medium (Cat: 11875-093, Gibco) containing 10% fetal bovine serum (BISH5400, BI), 1% penicillin/streptomycin, 50 uM 2-mercaptoethanol and were maintained in a humidified incubator with 5% CO2 at 37°C overnight. The cells were treated in 96-well culture plates (2.5 × 105 cells in 300 ul per well) with plate-bound anti-CD3 (2 μg/ml, immobilized overnight at 4°C) and fluid phase anti-CD28 (2 μg/ml), in the presence or absence of CRP (100 μg/ml), and then collected after 24 h for mRNA detection and 72 h for protein detection.
6
Isolation of Memory B Cells and Breg Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMCs were stained with CD19, CD24, and CD38 antibodies, and then CD19 + CD24 + CD38-memory B cells and CD19 + CD24hiCD38hi Breg cells were sorted with a MoFlo high-performance cell sorter (Cytomation, Fort Collins, CO, USA). CD4 + CD25- effector T cells were purified by magnetic-bead separation with MACS kits (Miltenyi, Gladbach, Germany) according to the manufacturer’s instructions, achieving >95% purity.
7
Generating Effector CD8+ T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary wild-type and Batf−/− P14 CD8+ T cells were isolated using CD8 negative selection magnetic-activated cell sorting (MACS) kits (Miltenyi) and cultured in RPMI 1640 supplemented with 10% fetal bovine serum, 10 mM HEPES, 50 U/ml of Penicillin, 50 μg/ml of Streptomycin, and 50 μM β-mercaptoethanol. Naive cells were stimulated with plate-bound anti-CD3 (4 μg/ml, clone 2C11, BD Pharmingen) and anti-CD28 (4 μg/ml, clone 37.51, BD Pharmingen) in the presence of recombinant human IL-2 (100 U/ml, R&D Systems) for 3 days to generate in vitro effector cells.
8
Enrichment and Depletion of T Cell Populations
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
OT-II cells used for in vitro co-culture and in vivo transfer experiments were enriched from the spleens of naïve OT-II mice using a negative selection “naïve CD4+ T cell enrichment kit” (Miltenyi, Germany). Purity was consistently between 88 and 96% CD4+ cells, with >99% of these cells being CD44−, as confirmed by flow cytometry.
CD3+, and or CD19+ cells were sometimes depleted from LN single cell suspensions using positive selection magnetic bead-based cell enrichments kits MACS kits (Miltenyi) as described in the manufacturer protocol.
CD3+, and or CD19+ cells were sometimes depleted from LN single cell suspensions using positive selection magnetic bead-based cell enrichments kits MACS kits (Miltenyi) as described in the manufacturer protocol.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!