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Pan t cells

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Pan-T cells are a type of immune cell used in research and clinical applications. They are a subset of T cells that express the CD3 cell surface marker, which is present on all mature T cells. Pan-T cells serve as a tool for the identification and isolation of T cells from various samples.

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Lab products found in correlation

Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Hek293t cell, by American Type Culture Collection (2 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (2 mentions) Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions) Fetal calf serum (fcs), by Gemini Bio (2 mentions) Negative selection, by Miltenyi Biotec (2 mentions) Jurkat t, by American Type Culture Collection (2 mentions) Cell proliferation reagent wst 1, by Merck Group (1 mentions) Cytotox 96 ldh substrate, by Promega (1 mentions) Anti cd25 and anti cd69 antibodies, by BD (1 mentions) Pan t cells, by AllCells (1 mentions) T cell activation expansion kit, by Miltenyi Biotec (1 mentions)

Spelling variants of this product

Pan T-cells isolation kit (6 citations) Pan T cells isolation kit II (4 citations) Human Pan T cells isolation kit (3 citations)

4 protocols using pan t cells

1

Cell culture and T cell activation

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HEK293T cells (human embryonic kidney cells expressing SV40 large T antigen, negative for mycoplasma, originally obtained from the American Type Culture Collection) and skin fibroblast cells derived from TNFAIP3-deficient patients or normal donors were grown in Dulbecco’s modified Eagle’s medium (Life Technologies) plus 10% fetal calf serum and 1x antibiotics (Life Technologies). Jurkat-T (human leukaemic T lymphoma cell line, negative for mycoplasma, originally obtained from the American Type Culture Collection) cells and human PBMCs were grown in RPMI1640 medium (Life Technologies), plus 10% fetal calf serum (Gemini Bio-Products) and antibiotics. For intracellular TNF staining, pan-T cells purified from PBMCs by negative selection (Miltenyi) were cultured in complete RPMI 1640 in the presence of plate-bound anti-CD3 and soluble anti-CD28 (both 1 μg/mL) for 5 days. For IL-9 staining, rhIL-4 (30 ng/mL), TGF-β (5 ng/mL), IL-1β (10 ng/ml), and IL-2 (10 U/ml), together with anti-IFNγ (10 μg/mL) were added to total PBMCs, and cells were cultured for 48 hours under the same conditions.
Zhou Q., Wang H., Schwartz D.M., Stoffels M., Park Y.H., Zhang Y., Yang D., Demirkaya E., Takeuchi M., Tsai W.L., Lyons J.J., Yu X., Ouyang C., Chen C., Chin D.T., Zaal K., Chandrasekharappa S.C., Hanson E.P., Yu Z., Mullikin J.C., Hasni S.A., Wertz I., Ombrello A.K., Stone D.L., Hoffmann P., Jones A., Barham B.K., Leavis H.L., van Royen-Kerkof A., Sibley C., Batu E.D., Gül A., Siegel R.M., Boehm M., Milner J.D., Ozen S., Gadina M., Chae J., Laxer R.M., Kastner D.L, & Aksentijevich I. (2015). Loss-of-function mutations in TNFAIP3 leading to A20 haploinsufficiency cause an early onset autoinflammatory syndrome. Nature genetics, 48(1), 67-73.
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2

In Vitro T-Cell Cytotoxicity Assay

Cited 1 time
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Target cells were plated at a density of 0.1 × 106 to 0.2 × 106 in RPMI-1640 + 10% FBS + 1% P/S, and frozen PBMCs (STEMCELL Technologies) were thawed, washed, and rested overnight. After 24 hours, pan T-cells were isolated (Miltenyi Biotec). pan T-cells were added to tumor cells at the E:T ratios indicated along with diluted antibody. After 48 hours, an aliquot of supernatant was removed, frozen, and subsequently analyzed for IL-2 and IFNγ release (MSD U-plex platform). Cytotoxicity was measured using Cell Proliferation Reagent WST-1 (Sigma) or CytoTox 96 LDH substrate (Promega). Percentage of cell lysis was calculated as previously described.25 (link)
Avanzino B.C., Prabhakar K., Dalvi P., Hartstein S., Kehm H., Balasubramani A., Boudreau A.A., Buelow B., Chang K., Davison L.M., Iyer S., Kalwit V., Lewis Wilson K., Malik-Chaudhry H.K., Pierson W., Pineda G., Rangaswamy U.S., Saiganesh S., Schellenberger U., Ugamraj H.S., Yabut R.D., Buelow R., Chapman J., Trinklein N.D, & Harris K.E. (2022). A T-cell engaging bispecific antibody with a tumor-selective bivalent folate receptor alpha binding arm for the treatment of ovarian cancer. Oncoimmunology, 11(1), 2113697.
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3

Cell culture and T cell activation

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HEK293T cells (human embryonic kidney cells expressing SV40 large T antigen, negative for mycoplasma, originally obtained from the American Type Culture Collection) and skin fibroblast cells derived from TNFAIP3-deficient patients or normal donors were grown in Dulbecco’s modified Eagle’s medium (Life Technologies) plus 10% fetal calf serum and 1x antibiotics (Life Technologies). Jurkat-T (human leukaemic T lymphoma cell line, negative for mycoplasma, originally obtained from the American Type Culture Collection) cells and human PBMCs were grown in RPMI1640 medium (Life Technologies), plus 10% fetal calf serum (Gemini Bio-Products) and antibiotics. For intracellular TNF staining, pan-T cells purified from PBMCs by negative selection (Miltenyi) were cultured in complete RPMI 1640 in the presence of plate-bound anti-CD3 and soluble anti-CD28 (both 1 μg/mL) for 5 days. For IL-9 staining, rhIL-4 (30 ng/mL), TGF-β (5 ng/mL), IL-1β (10 ng/ml), and IL-2 (10 U/ml), together with anti-IFNγ (10 μg/mL) were added to total PBMCs, and cells were cultured for 48 hours under the same conditions.
Zhou Q., Wang H., Schwartz D.M., Stoffels M., Park Y.H., Zhang Y., Yang D., Demirkaya E., Takeuchi M., Tsai W.L., Lyons J.J., Yu X., Ouyang C., Chen C., Chin D.T., Zaal K., Chandrasekharappa S.C., Hanson E.P., Yu Z., Mullikin J.C., Hasni S.A., Wertz I., Ombrello A.K., Stone D.L., Hoffmann P., Jones A., Barham B.K., Leavis H.L., van Royen-Kerkof A., Sibley C., Batu E.D., Gül A., Siegel R.M., Boehm M., Milner J.D., Ozen S., Gadina M., Chae J., Laxer R.M., Kastner D.L, & Aksentijevich I. (2015). Loss-of-function mutations in TNFAIP3 leading to A20 haploinsufficiency cause an early onset autoinflammatory syndrome. Nature genetics, 48(1), 67-73.
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4

T Cell Activation Methods for Assays

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Pan-T cells were purchased from AllCells. T cells that were not stimulated prior to use in assays are referred to as resting T cells. Activated T cells were generated by two methods: (1) for in vivo studies, activated T cells were generated using beads coated with anti-CD2, -CD28 and -CD3 antibodies for 3–7 days following the manufacturer’s instructions (Miltenyi Biotec T cell activation/expansion kit); (2) for in vitro assays, BiTE®-activated T cells were generated in bulk by culturing Pan-T cells with NUGC4 cells (10:1) and 100 pM EGFR BiTE® for 24 hours prior to use in assays. T cells were washed with fresh media before using in assays. Activation was confirmed using anti-CD25 and anti-CD69 antibodies (BD).
Ross S.L., Sherman M., McElroy P.L., Lofgren J.A., Moody G., Baeuerle P.A., Coxon A, & Arvedson T. (2017). Bispecific T cell engager (BiTE®) antibody constructs can mediate bystander tumor cell killing. PLoS ONE, 12(8), e0183390.
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