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αcd3 αcd28 beads

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The αCD3/αCD28 beads are a type of magnetic beads coated with antibodies against the CD3 and CD28 proteins. These beads are commonly used in cell culture and cell activation applications to stimulate and expand T cells.

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Lab products found in correlation

Cd8 t cell isolation kit, by Miltenyi Biotec (2 mentions) Automacs, by Miltenyi Biotec (2 mentions) Ficoll purified pbmc, by Miltenyi Biotec (2 mentions) Cgmp grade anti cd25 microbeads, by Miltenyi Biotec (2 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Recombinant murine il 2, by Thermo Fisher Scientific (1 mentions) Negative selection magnetic beads, by Miltenyi Biotec (1 mentions) Protein transport inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Retronectin, by Takara Bio (1 mentions)

Spelling variants of this product

αCD3/αCD28 activating beads (4 citations) αCD3/αCD28 antibody-conjugated beads (3 citations)

4 protocols using αcd3 αcd28 beads

1

Regulation of CD4+ T Cell Activation by EV

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Naive splenic CD4+ T lymphocytes were captured using negative selection magnetic beads (Miltenyi Biotec, Germany). These cells were stimulated with αCD3/αCD28 beads (Thermo Fisher, USA) and 5 ng/ml of recombinant murine IL‐2 (Peprotech, USA) for 48 h while also being co‐cultured with 2×109 particles/mL of EKO‐BMDM‐EV, or WT‐BMDM‐EV, or PBS. Measurements of T lymphocyte activation were assessed using flow cytometric detection of CD69 and CD25. For detection of IFN‐γ, naive splenic CD4+ T lymphocytes were stimulated with αCD3/αCD28 beads (Thermo Fisher, USA) and 5 ng/ml of recombinant murine IL‐2 (Peprotech, USA) for 12 h while also being co‐cultured with 2×109 particles/mL of EKO‐BMDM‐EV, or WT‐BMDM‐EV, or PBS. Cells were cultured in the presence of the Protein Transport Inhibitor cocktail (Invitrogen, USA). They were then permeabilized using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience, USA) and stained with anti‐IFNγ (clone XMG1.2) at 1:100 concentration for 60 min in room temperature according to the manufacturer's protocol.
Phu T.A., Ng M., Vu N.K., Gao A.S, & Raffai R.L. (2023). ApoE expression in macrophages communicates immunometabolic signaling that controls hyperlipidemia‐driven hematopoiesis & inflammation via extracellular vesicles. Journal of Extracellular Vesicles, 12(8), 12345.
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2

Retroviral Transduction of GD2-CAR T Cells

Cited 1 time
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Retroviral supernatant for the third generation GD2-CAR (SGF.iCasp9.2A.14g2a.CD28.OX40.ζ) was harvested from the producer cell clone. Retroviral supernatants of the previously described GD2-specific CARs incorporating either CD28 or 4-1BB (MSGV.14g2a.CD28.ζ or MSGV.14g2a.4-1BB.ζ) were produced by transient transfection of the 293GP cell line using the RD114 envelope protein (15 (link)). Cryopreserved PBMCs were thawed and activated using αCD3/αCD28 beads (Invitrogen) at a 3:1 bead:T-cell ratio with 40 IU/mL rhIL2 for 3 days. Non-tissue coated 6-well plates were coated with 24 ug/well retronectin (Takara) for 2 hours at room temperature, and then blocked with 2.5% BSA for 30 min at room temperature. Plates were spin-coated with retroviral supernatant at 3050 rpm at 32°C for 3 hours. Activated human lymphocytes were added at 106 cells per well. Transduction was repeated the following day and cells were expanded in AIM-V media with 300 IU/mL rhIL2.
Long A.H., Highfill S.L., Cui Y., Smith J.P., Walker A.J., Ramakrishna S., El-Etriby R., Galli S., Tsokos M.G., Orentas R.J, & Mackall C.L. (2016). Reduction of MDSCs with all-trans retinoic acid improves CAR therapy efficacy for sarcomas. Cancer immunology research, 4(10), 869-880.
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3

Isolation and Stimulation of Resting CD8+ T Cells

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Resting CD8+ T cells were isolated from Ficoll-purified PBMC isolated from leukapheresis products (Memorial Blood Center, St. Paul, MN) in a two-step process by first depleting CD25+ cells using using (cGMP)-grade anti-CD25 microbeads (Miltenyi Biotec, Auburn, CA) on an AutoMACS (Miltenyi Biotec). CD8+ cells were then purified from the CD25− fraction using negative selection (CD8+ T cell isolation kit, Miltenyi Biotec). Purified cells were stimulated for 72 h with αCD3/αCD28 beads (InVitrogen) + IL-2 (300U/ml). After 20 h, A-438079 (25µM) or PBS was added to the cultures. Cell numbers were analyzed by Neubauer chamber counting. Survival was analyzed by Trypan blue staining during microscope counting and confirmed by Live-Dead (Tonbo Biosciences) staining using flow cytometry.
da Silva H.B., Beura L.K., Wang H., Hanse E.A., Gore R., Scott M.C., Walsh D.A., Block K.E., Fonseca R., Yan Y., Hippen K.L., Blazar B.R., Masopust D., Kelekar A., Vulchanova L., Hogquist K.A, & Jameson S.C. (2018). The purinergic receptor P2RX7 directs metabolic fitness of long-lived memory CD8+ T cells. Nature, 559(7713), 264-268.
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4

Isolation and Stimulation of Resting CD8+ T Cells

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Resting CD8+ T cells were isolated from Ficoll-purified PBMC isolated from leukapheresis products (Memorial Blood Center, St. Paul, MN) in a two-step process by first depleting CD25+ cells using using (cGMP)-grade anti-CD25 microbeads (Miltenyi Biotec, Auburn, CA) on an AutoMACS (Miltenyi Biotec). CD8+ cells were then purified from the CD25− fraction using negative selection (CD8+ T cell isolation kit, Miltenyi Biotec). Purified cells were stimulated for 72 h with αCD3/αCD28 beads (InVitrogen) + IL-2 (300U/ml). After 20 h, A-438079 (25µM) or PBS was added to the cultures. Cell numbers were analyzed by Neubauer chamber counting. Survival was analyzed by Trypan blue staining during microscope counting and confirmed by Live-Dead (Tonbo Biosciences) staining using flow cytometry.
da Silva H.B., Beura L.K., Wang H., Hanse E.A., Gore R., Scott M.C., Walsh D.A., Block K.E., Fonseca R., Yan Y., Hippen K.L., Blazar B.R., Masopust D., Kelekar A., Vulchanova L., Hogquist K.A, & Jameson S.C. (2018). The purinergic receptor P2RX7 directs metabolic fitness of long-lived memory CD8+ T cells. Nature, 559(7713), 264-268.
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