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Mojosort human cd3 t cell isolation kit

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The MojoSort Human CD3 T Cell Isolation Kit is a magnetic-based isolation kit designed to enrich for human CD3+ T cells from a variety of sample types. The kit utilizes a negative selection approach to isolate the target cells without directly labeling them, allowing for the recovery of highly pure and unactivated T cells.

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Lab products found in correlation

Cd3 coated plates clone okt3, by BioLegend (2 mentions) Easysep human cd8 t cell isolation kit, by STEMCELL (2 mentions) Cd8 microbead, by Miltenyi Biotec (2 mentions) Mojosort human cd8 t cell isolation kit, by BioLegend (2 mentions) Dynabead, by Thermo Fisher Scientific (2 mentions) Ficoll hypaque, by Merck Group (1 mentions) Dynabeads human t activator cd3 cd28, by Thermo Fisher Scientific (1 mentions) Dynabeads human t activator cd3 cd28 for t cell expansion and activation, by Thermo Fisher Scientific (1 mentions) Histopaque 1077, by Merck Group (1 mentions) Fixation permeabilization solution kit, by BD (1 mentions) Fortessa, by BD (1 mentions) Lsr 2, by BD (1 mentions) Ultracomp ebeads, by Thermo Fisher Scientific (1 mentions) Fcs express, by De Novo Software (1 mentions) Foxp3 transcription factor staining buffer, by Thermo Fisher Scientific (1 mentions) Influx high speed sorter, by BD (1 mentions) Ficoll paque plus, by Merck Group (1 mentions) Cell activation cocktail, by BioLegend (1 mentions) Tag it violet proliferation and cell tracking dye, by BioLegend (1 mentions) Celltiter blue cell viability assay, by Promega (1 mentions)

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Human CD3 T cell isolation kit (3 citations) Mojosort human CD3 isolation kit (2 citations)

19 protocols using mojosort human cd3 t cell isolation kit

1

CD69 Expression in CD3+ T Cells

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Peripheral blood from normal, healthy donors was collected into potassium EDTA vacates. Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll® lymphocyte separation medium (LSM, MP Biomedicals, Aurora, USA) centrifugation as described previously (Chikamatsu et al, 2007). Next, CD3 T cells were separated from freshly PBMCs by using a MojoSortTM Human CD3+ T cell Isolation Kit (BioLegend, https://www.biolegend.com/protocols/mojosort‐isolation‐kits‐protocol‐1/4599/).
For CD69 early activation detection, freshly CD3+ T cells were infected with shNC, shNF45 or shNF90 lentivirus; then, cells were cultured in plates coated with anti‐CD3 and ‐CD28 mAbs and stimulated with P/I for 12 h. The CD3‐positive T cells were further pre‐gated with anti‐CD3 antibody, and CD69 expression was analyzed using an anti‐CD69 antibody by FACS.
Tsai H., Zeng X., Liu L., Xin S., Wu Y., Xu Z., Zhang H., Liu G., Bi Z., Su D., Yang M., Tao Y., Wang C., Zhao J., Eriksson J.E., Deng W., Cheng F, & Chen H. (2021). NF45/NF90‐mediated rDNA transcription provides a novel target for immunosuppressant development. EMBO Molecular Medicine, 13(3), e12834.
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2

ADAM17 Inhibition in Activated CD3+ T Cells

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CD3+ T cells were isolated from PBMCs using a MojoSortTM Human CD3 T Cell Isolation Kit (BioLegend, USA). The purity of isolated CD3+ T cells was above 95%, as examined by flow cytometry. ADAM17 inhibition experiment was carried out according to previous report [41 (link)]. Isolated CD3+ T cells were cultured in X–VIVOTM 15 medium containing 20% FBS, and treated for 1 h in the presence or absence of TAPI-1 (20 mM). Next, they were incubated on a plate coated with anti-CD3 antibody (10 μg/ml) and anti-CD28 antibody (1 μg/ml) for 4 h. Cells were harvested after treatment, and flow cytometry was performed to measure the fluorescence intensity of ADAM17 and IL-23 R in CD4+ T cells.
Wang X., Xin S., Wang Y., Ju D., Wu Q., Qiu Y., Niu X., Liu W., Li J, & Ji P. (2021). MicroRNA-146a-5p enhances T helper 17 cell differentiation via decreasing a disintegrin and metalloprotease 17 level in primary sjögren’s syndrome. Bioengineered, 12(1), 310-324.
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3

Isolation of Highly Purified CD3+ T Cells

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Peripheral blood from normal healthy donors was collected into EDTA potassium vacuoles. PBMCs were isolated using Ficoll lymphocyte isolation medium. Next, CD3+ T lymphocytes were purified (>98%) by negative selection with the MojoSortTM Human CD3+ T cell Isolation Kit (Biolegend, USA). Cells were seeded on CD3‐coated plates (clone OKT3; Biolegend) and cultured in medium (RPMI1640 with 10% FBS, penicillin/streptomycin, and 2 ng ml–1 Il‐2).
Tsai H., Wu Y., Liu X., Xu Z., Liu L., Wang C., Zhang H., Huang Y., Wang L., Zhang W., Su D., Khan F.U., Zhu X., Yang R., Pang Y., Eriksson J.E., Zhu H., Wang D., Jia B., Cheng F, & Chen H. (2021). Engineered Small Extracellular Vesicles as a FGL1/PD‐L1 Dual‐Targeting Delivery System for Alleviating Immune Rejection. Advanced Science, 9(3), 2102634.
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4

Isolation and Culture of Human CD3+ T Cells

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Peripheral blood from normal healthy donors was collected into EDTA potassium vacuoles.
PBMCs were isolated using Ficoll lymphocyte isolation medium. Next, CD3 + T lymphocytes were purified (> 98%) by negative selection with MojoSort TM Human CD3 + T cell Isolation Kit (Biolegend, USA). Cells were seeded in CD3-coated plates (clone OKT3; Biolegend) and cultured in medium (RPMI1640 with 10 % FBS, penicillin/streptomycin, and 2 ng mL -1 IL-2).
Tsai H., Wu Y., Liu X., Xu Z., Liu L., Wang C., Zhang H., Huang Y., Wang L., Zhang W., Su D., Khan F.U., Zhu X., Yang R., Pang Y., Eriksson J., Zhu H., Wang D., Jia B., Cheng F., & Chen H. (2021). Engineered small extracellular vesicles as a FGL1/PD-L1 dual-targeting delivery system for alleviating immune rejection.
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5

PBMC Isolation, T-cell Enrichment, and NALM-6 Transduction

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Peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats via density gradient separation and either used fresh or cryopreserved in fetal bovine serum (FBS) supplemented with 10% dimethyl sulfoxide (DMSO). Tissue samples were processed as described previously16 (link),51 (link),52 (link). Total CD8+ T cells were enriched via negative magnetic separation using an EasySep Human CD8+ T Cell Isolation Kit (Stem Cell Technologies) or a MojoSort Human CD8+ T Cell Isolation Kit (BioLegend). Total CD3+ T cells were enriched via negative magnetic separation using a MojoSort Human CD3+ T Cell Isolation Kit (BioLegend). CD8-depleted PBMCs were obtained via negative magnetic separation using CD8 MicroBeads (Miltenyi Biotec). The human NALM-6 cell line (DSMZ) was tested for Mycoplasma (Eurofins Genomics) and transduced with a lentiviral vector encoding secreted luciferase (Lucia+/NGFR+/NALM-6)53 (link).
Galletti G., De Simone G., Mazza E.M., Puccio S., Mezzanotte C., Bi T.M., Davydov A.N., Metsger M., Scamardella E., Alvisi G., De Paoli F., Zanon V., Scarpa A., Camisa B., Colombo F.S., Anselmo A., Peano C., Polletti S., Mavilio D., Gattinoni L., Boi S.K., Youngblood B.A., Jones R.E., Baird D.M., Gostick E., Llewellyn-Lacey S., Ladell K., Price D.A., Chudakov D.M., Newell E.W., Casucci M, & Lugli E. (2020). Two subsets of stem-like CD8+ memory T cell progenitors with distinct fate commitments in humans. Nature immunology, 21(12), 1552-1562.
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6

Isolating T and B Cells from Monkey PBMCs

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Peripheral blood mononuclear cells (PBMC) from monkeys were isolated by standard density gradient centrifugation (Ficoll‐Hypaque, Sigma Chemicals®). Then T cells were harvested using MojoSort™ Human CD3 T cell isolation kit (Biolegend) and B cells were isolated using EasySep™ Human B Cells Enrichment Kit (Stem Cell Technology).
Guo G., Zhu Y., Wu Z., Ji H., Lu X., Zhou Y., Li Y., Cao X., Lu Y., Talbot P., Liao J., Shi Y, & Bu H. (2018). Circulating monocytes accelerate acute liver failure by IL‐6 secretion in monkey. Journal of Cellular and Molecular Medicine, 22(9), 4056-4067.
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7

PBMC Isolation, T-cell Enrichment, and NALM-6 Transduction

Cited 1 time
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Peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats via density gradient separation and either used fresh or cryopreserved in fetal bovine serum (FBS) supplemented with 10% dimethyl sulfoxide (DMSO). Tissue samples were processed as described previously16 (link),51 (link),52 (link). Total CD8+ T cells were enriched via negative magnetic separation using an EasySep Human CD8+ T Cell Isolation Kit (Stem Cell Technologies) or a MojoSort Human CD8+ T Cell Isolation Kit (BioLegend). Total CD3+ T cells were enriched via negative magnetic separation using a MojoSort Human CD3+ T Cell Isolation Kit (BioLegend). CD8-depleted PBMCs were obtained via negative magnetic separation using CD8 MicroBeads (Miltenyi Biotec). The human NALM-6 cell line (DSMZ) was tested for Mycoplasma (Eurofins Genomics) and transduced with a lentiviral vector encoding secreted luciferase (Lucia+/NGFR+/NALM-6)53 (link).
Galletti G., De Simone G., Mazza E.M., Puccio S., Mezzanotte C., Bi T.M., Davydov A.N., Metsger M., Scamardella E., Alvisi G., De Paoli F., Zanon V., Scarpa A., Camisa B., Colombo F.S., Anselmo A., Peano C., Polletti S., Mavilio D., Gattinoni L., Boi S.K., Youngblood B.A., Jones R.E., Baird D.M., Gostick E., Llewellyn-Lacey S., Ladell K., Price D.A., Chudakov D.M., Newell E.W., Casucci M, & Lugli E. (2020). Two subsets of stem-like CD8+ memory T cell progenitors with distinct fate commitments in humans. Nature immunology, 21(12), 1552-1562.
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8

Isolation and Activation of Human T Cells

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In this study, peripheral blood mononuclear cells (PBMCs) were retrieved from the venous blood of healthy volunteers using Ficoll density gradient centrifugation. To isolate T cells, a specific number of PBMCs were resuspended in 100 µL of separation buffer and subjected to sorting using the MojoSort Human CD3 T Cell Isolation Kit (BioLegend; Cat. no. 480022). To sort the CD3 + T cells, 10 µL of CD3 negative antibody was added to each 1 × 107 cells. Once sorted, the cells were transferred to X-VIVO15 medium (serum-free hematopoietic cell medium) supplemented with 5% human serum for T cell culture. To activate the CD3 + T cells, CD3/CD28 magnetic beads (Dynabeads Human T-activator CD3/CD28, Life Technologies, Cat. no. 11131D) were employed. The activated CD3 + T cells were then utilized in experiments conducted a week later.
Wang S.Y., Zhang S.J., Meng H.F., Xu H.Q., Guo Z.X., Yan J.F., Gao J.L., Niu L.N., Wang S.L, & Jiao K. (2024). DPSCs regulate epithelial-T cell interactions in oral submucous fibrosis. Stem Cell Research & Therapy, 15, 113.
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9

Activated T cell co-culture with BxPC-3 cells

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CD3 T lymphocyte isolation was performed using the isolation protocol from Biolegend (MojoSort™ Human CD3 T Cell Isolation Kit, Catalog #480131, San Diego, CA, USA). T cell activation and proliferation were performed according to the manufacturer’s protocol of Dynabeads™ Human T-Activator CD3/CD28 for T Cell Expansion and Activation (Catalog#11131D, Thermo Fisher Scientific, Waltham, MA, USA). Coculture of activated T cells with BxPC-3 used a cell ratio of 1:1 in RPMI 1640 supplemented with 100 U/mL penicillin, 100 mg/mL streptomycin, 1 mM L-glutamine, and 10% FBS in the presence of CF33-hNIS-antiPDL1 (MOI = 3) for 5 days.
Zhang Z., Yang A., Chaurasiya S., Park A.K., Lu J., Kim S.I., Warner S.G., Yuan Y.C., Liu Z., Han H., Von Hoff D., Fong Y, & Woo Y. (2021). CF33-hNIS-antiPDL1 Virus Primes Pancreatic Ductal Adenocarcinoma for Enhanced anti-PD-L1 Therapy. Cancer gene therapy, 29(6), 722-733.
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10

Isolation of Untouched CD3+ T Cells from Healthy Donors

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Healthy donors’ datasets were downloaded from the Gene Expression Omnibus (GEO, accession number: GSE126030) (12 (link)). The samples were obtained from deceased, brain-dead donors at the time of organ acquisition for clinical transplantation. Donors were free of chronic disease, cancer, and chronic infections such as Hepatitis B, C, and HIV. The mononuclear cells were isolated from human lungs (LG), lymph nodes (LN), bone marrow (BM), and blood, and the untouched CD3+ T cells were enriched from single-cell suspensions of all tissues and blood using magnetic negative selection (MojoSort Human CD3+ T cell Isolation Kit; BioLegend) (12 (link)).
Li Z., Wang H., Dong R., Man J., Sun L., Qian X., Zhu X., Cao P., Yu Y., Le J., Fu Y., Wang P., Jiang W., Shen C., Ma Y., Chen L., Xu Y., Shi J., Zhang H., Qian M, & Zhai X. (2021). Single-Cell RNA-seq Reveals Characteristics of Malignant Cells and Immune Microenvironment in Subcutaneous Panniculitis-Like T-Cell Lymphoma. Frontiers in Oncology, 11, 611580.
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