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Anti cd3 clone okt3

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Anti-CD3 (clone OKT3) is a monoclonal antibody that specifically binds to the CD3 complex on the surface of T cells. It is used in research applications to activate and study T cell function.

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Lab products found in correlation

Anti cd28 clone cd28, by Thermo Fisher Scientific (9 mentions) Rosiglitazone, by Merck Group (6 mentions) Anti cd28 clone cd28, by BD (4 mentions) T cell purification kit, by Thermo Fisher Scientific (4 mentions) Reverse transcription master mix, by Standard BioTools (3 mentions) Preamp master mix, by Standard BioTools (3 mentions) Evagreen supermix, by Bio-Rad (3 mentions) Biomark hd system, by Standard BioTools (3 mentions) Nucleospin rna xs kit, by Macherey-Nagel (3 mentions) Cell proliferation dye efluor 450, by Thermo Fisher Scientific (3 mentions) Aim 5 medium, by Thermo Fisher Scientific (3 mentions) Anti cd69 fitc clone l78, by BD (3 mentions) Anti cd134 bv711 clone act35, by BD (3 mentions) Anti active caspase 3 pe clone c92 605, by BD (3 mentions) Fenofibrate, by Merck Group (3 mentions) Cytofix cytoperm fixation permeabilization solution kit, by BD (3 mentions) Facscalibur, by BD (2 mentions) Recombinant human il 2, by R&D Systems (2 mentions) Cdh26 higg1 fc, by R&D Systems (2 mentions) Cdh1 fc, by R&D Systems (2 mentions)

47 protocols using anti cd3 clone okt3

1

Activated CD8+ T Cell Gene Expression

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PBMCs were activated for 5 hr with plate-bound anti-CD3 (clone OKT3; Thermo Fisher Scientific). RNA was extracted from flow-sorted naive CD8+ T cells (n = 300 per condition) using a NucleoSpin RNA XS Kit (Macherey-Nagel), and cDNA was synthesized using Reverse Transcription Master Mix (Fluidigm). Specific targets were amplified using PreAmp Master Mix (Fluidigm). Gene expression was assessed using a BioMark HD System (Fluidigm) with EvaGreen Supermix (Bio-Rad). RNA expression levels were calculated using the 2−ΔΔCT method with reference to a housekeeping gene (human 18S) (18 (link)).
Nicoli F., Cabral-Piccin M.P., Papagno L., Gallerani E., Fusaro M., Folcher V., Dubois M., Clave E., Vallet H., Frere J.J., Gostick E., Llewellyn-Lacey S., Price D.A., Toubert A., Dupré L., Boddaert J., Caputo A., Gavioli R, & Appay V. (2022). Altered basal lipid metabolism underlies the functional impairment of naive CD8+ T cells in elderly humans. Journal of immunology (Baltimore, Md. : 1950), 208(3), 562-570.
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2

T-cell Proliferation Modulation by Fatty Acids

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PBMCs were labeled with Cell Proliferation Dye (CPD) eFluor450 (Thermo Fisher Scientific) and stimulated for 4 days with plate-bound anti-CD3 (clone OKT3; Thermo Fisher Scientific). In some experiments, cells were precultured in AIM-V medium (Thermo Fisher Scientific) supplemented with BSA (10% v/v; Sigma-Aldrich) for 1 day in the absence or presence of palmitic acid (PA; 300 μM; Sigma-Aldrich), and in other experiments, cells were precultured in AIM-V medium (Thermo Fisher Scientific) without BSA supplementation for 2 days in the absence or presence of rosiglitazone (40 μM; Sigma-Aldrich). Proliferation was measured using flow cytometry to quantify the dilution of CPD. Sample size calculation, based on available data for T-cell proliferation (5 (link)), suggested a size of 10 individuals per group to detect a 50% difference in CPD low (i.e. proliferating) cells between middle-aged and old individuals with a power of 80% using a one-sided significance level of 5%. To assess the effect of fatty acids on resting cells, PBMCs were cultured up to 48 hr in AIM-V medium supplemented with 10% BSA in the absence or in the presence of palmitic acid (300 or 3000 μM).
Nicoli F., Cabral-Piccin M.P., Papagno L., Gallerani E., Fusaro M., Folcher V., Dubois M., Clave E., Vallet H., Frere J.J., Gostick E., Llewellyn-Lacey S., Price D.A., Toubert A., Dupré L., Boddaert J., Caputo A., Gavioli R, & Appay V. (2022). Altered basal lipid metabolism underlies the functional impairment of naive CD8+ T cells in elderly humans. Journal of immunology (Baltimore, Md. : 1950), 208(3), 562-570.
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3

Activated CD8+ T Cell Gene Expression

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PBMCs were activated for 5 hr with plate-bound anti-CD3 (clone OKT3; Thermo Fisher Scientific). RNA was extracted from flow-sorted naive CD8+ T cells (n = 300 per condition) using a NucleoSpin RNA XS Kit (Macherey-Nagel), and cDNA was synthesized using Reverse Transcription Master Mix (Fluidigm). Specific targets were amplified using PreAmp Master Mix (Fluidigm). Gene expression was assessed using a BioMark HD System (Fluidigm) with EvaGreen Supermix (Bio-Rad). RNA expression levels were calculated using the 2−ΔΔCT method with reference to a housekeeping gene (human 18S) (18 (link)).
Nicoli F., Cabral-Piccin M.P., Papagno L., Gallerani E., Fusaro M., Folcher V., Dubois M., Clave E., Vallet H., Frere J.J., Gostick E., Llewellyn-Lacey S., Price D.A., Toubert A., Dupré L., Boddaert J., Caputo A., Gavioli R, & Appay V. (2022). Altered basal lipid metabolism underlies the functional impairment of naive CD8+ T cells in elderly humans. Journal of immunology (Baltimore, Md. : 1950), 208(3), 562-570.
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4

Anti-CD3 Stimulation and PPAR Modulation

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PBMCs were stimulated for 24 hr with plate-bound anti-CD3 (clone OKT3; Thermo Fisher Scientific) in the absence or presence of fenofibrate (50 μM; Sigma-Aldrich) or rosiglitazone (40 μM; Sigma-Aldrich). Activation markers were quantified on the cell surface using anti-CD69–FITC (clone L78; BD Biosciences) and anti-CD134–BV711 (clone ACT35; BD Biosciences). Intracellular stains were performed using anti-active caspase-3–PE (clone C92-605; BD Biosciences) in conjunction with a Cytofix/Cytoperm Fixation/Permeabilization Solution Kit (BD Biosciences).
Nicoli F., Cabral-Piccin M.P., Papagno L., Gallerani E., Fusaro M., Folcher V., Dubois M., Clave E., Vallet H., Frere J.J., Gostick E., Llewellyn-Lacey S., Price D.A., Toubert A., Dupré L., Boddaert J., Caputo A., Gavioli R, & Appay V. (2022). Altered basal lipid metabolism underlies the functional impairment of naive CD8+ T cells in elderly humans. Journal of immunology (Baltimore, Md. : 1950), 208(3), 562-570.
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5

Anti-CD3 Stimulation and PPAR Modulation

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PBMCs were stimulated for 24 hr with plate-bound anti-CD3 (clone OKT3; Thermo Fisher Scientific) in the absence or presence of fenofibrate (50 μM; Sigma-Aldrich) or rosiglitazone (40 μM; Sigma-Aldrich). Activation markers were quantified on the cell surface using anti-CD69–FITC (clone L78; BD Biosciences) and anti-CD134–BV711 (clone ACT35; BD Biosciences). Intracellular stains were performed using anti-active caspase-3–PE (clone C92-605; BD Biosciences) in conjunction with a Cytofix/Cytoperm Fixation/Permeabilization Solution Kit (BD Biosciences).
Nicoli F., Cabral-Piccin M.P., Papagno L., Gallerani E., Fusaro M., Folcher V., Dubois M., Clave E., Vallet H., Frere J.J., Gostick E., Llewellyn-Lacey S., Price D.A., Toubert A., Dupré L., Boddaert J., Caputo A., Gavioli R, & Appay V. (2022). Altered basal lipid metabolism underlies the functional impairment of naive CD8+ T cells in elderly humans. Journal of immunology (Baltimore, Md. : 1950), 208(3), 562-570.
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6

Activated B Cell-CD4+ T Cell Co-culture

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B cells were isolated as described and cultured at a density of 1 × 106 cells per ml for 48 h with 1 μM CpGC, with and without inhibitors. After 48 h, B cells were washed and incubated 1:1 with autologous resting CD4+ T cells (T cells isolated using the EasySep Human CD4+ T cell Isolation Kit, STEMCELL, cat. no. 17962) for a further 72 h in culture plates coated with plate-bound anti-CD3 (clone OKT3, ThermoFisher Scientific, cat. no. 16-0037-81).
Bradford H.F., McDonnell T.C., Stewart A., Skelton A., Ng J., Baig Z., Fraternali F., Dunn-Walters D., Isenberg D.A., Khan A.R., Mauro C, & Mauri C. (2024). Thioredoxin is a metabolic rheostat controlling regulatory B cells. Nature Immunology, 25(5), 873-885.
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7

T-cell Proliferation Modulation by Fatty Acids

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
PBMCs were labeled with Cell Proliferation Dye (CPD) eFluor450 (Thermo Fisher Scientific) and stimulated for 4 days with plate-bound anti-CD3 (clone OKT3; Thermo Fisher Scientific). In some experiments, cells were precultured in AIM-V medium (Thermo Fisher Scientific) supplemented with BSA (10% v/v; Sigma-Aldrich) for 1 day in the absence or presence of palmitic acid (PA; 300 μM; Sigma-Aldrich), and in other experiments, cells were precultured in AIM-V medium (Thermo Fisher Scientific) without BSA supplementation for 2 days in the absence or presence of rosiglitazone (40 μM; Sigma-Aldrich). Proliferation was measured using flow cytometry to quantify the dilution of CPD. Sample size calculation, based on available data for T-cell proliferation (5 (link)), suggested a size of 10 individuals per group to detect a 50% difference in CPD low (i.e. proliferating) cells between middle-aged and old individuals with a power of 80% using a one-sided significance level of 5%. To assess the effect of fatty acids on resting cells, PBMCs were cultured up to 48 hr in AIM-V medium supplemented with 10% BSA in the absence or in the presence of palmitic acid (300 or 3000 μM).
Nicoli F., Cabral-Piccin M.P., Papagno L., Gallerani E., Fusaro M., Folcher V., Dubois M., Clave E., Vallet H., Frere J.J., Gostick E., Llewellyn-Lacey S., Price D.A., Toubert A., Dupré L., Boddaert J., Caputo A., Gavioli R, & Appay V. (2022). Altered basal lipid metabolism underlies the functional impairment of naive CD8+ T cells in elderly humans. Journal of immunology (Baltimore, Md. : 1950), 208(3), 562-570.
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8

Investigating T Cell Proliferation Modulation

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PBMCs were labeled with Cell Proliferation Dye (CPD) eFluor450 (Thermo Fisher Scientific) and stimulated for 4 days with plate-bound anti-CD3 (clone OKT3; Thermo Fisher Scientific) in the absence or presence of CTAB (1 μM; Sigma-Aldrich). In some experiments, cells were precultured in AIM-V medium (Thermo Fisher Scientific) supplemented with bovine serum albumin (BSA; 10% v/v; Sigma-Aldrich) for 1 day in the absence or presence of palmitic acid (300 μM; Sigma-Aldrich), and in other experiments, cells were precultured in AIM-V medium (Thermo Fisher Scientific) without BSA supplementation for 2 days in the absence or presence of rosiglitazone (40 μM; Sigma-Aldrich). Proliferation was measured using flow cytometry to quantify the dilution of CPD.
Nicoli F., Cabral-Piccin M.P., Papagno L., Gallerani E., Folcher V., Dubois M., Barrett A.J., Vallet H., Frere J.J., Gostick E., Llewellyn‐Lacey S., Price D.A., Toubert A., Boddaert J., Caputo A., Gavioli R., & Appay V. (2020). Altered basal lipid metabolism underlies the functional impairment of naive CD8+ T cells in elderly humans.
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9

Profiling Activated CD8+ T Cell Transcriptome

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PBMCs were activated for 5 hr with plate-bound anti-CD3 (clone OKT3; Thermo Fisher Scientific). RNA was extracted from flow-sorted naive CD8 + T cells (n = 300 per condition) using a NucleoSpin RNA XS Kit (Macherey-Nagel), and cDNA was synthesized using Reverse Transcription Master Mix (Fluidigm). Specific targets were amplified using PreAmp Master Mix (Fluidigm). Gene expression was assessed using a BioMark HD System (Fluidigm) with EvaGreen Supermix (Bio-Rad). RNA expression levels were calculated using the 2 -ΔΔCT method with reference to a housekeeping gene (human 18S) [16] .
Nicoli F., Cabral-Piccin M.P., Papagno L., Gallerani E., Folcher V., Dubois M., Barrett A.J., Vallet H., Frere J.J., Gostick E., Llewellyn‐Lacey S., Price D.A., Toubert A., Boddaert J., Caputo A., Gavioli R., & Appay V. (2020). Altered basal lipid metabolism underlies the functional impairment of naive CD8+ T cells in elderly humans.
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10

Activation Markers and Apoptosis in PBMCs

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PBMCs were stimulated for 24 hr with plate-bound anti-CD3 (clone OKT3; Thermo Fisher Scientific) in the absence or presence of fenofibrate (50 μM; Sigma-Aldrich) or rosiglitazone (40 Sigma-Aldrich). The expression of the standard activation markers CD69 and CD134 (OX40) was quantified on the cell surface using anti-CD69-FITC (clone L78; BD Biosciences) and anti-CD134-BV711 (clone ACT35; BD Biosciences). Intracellular staining for activated caspase-3, used as marker of susceptibility to apoptosis upon activation, was performed using anti-active caspase-3-PE (clone C92-605; BD Biosciences) in conjunction with a Cytofix/Cytoperm Fixation/Permeabilization Solution Kit (BD Biosciences).
Nicoli F., Cabral-Piccin M.P., Papagno L., Gallerani E., Folcher V., Dubois M., Barrett A.J., Vallet H., Frere J.J., Gostick E., Llewellyn‐Lacey S., Price D.A., Toubert A., Boddaert J., Caputo A., Gavioli R., & Appay V. (2020). Altered basal lipid metabolism underlies the functional impairment of naive CD8+ T cells in elderly humans.
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