B6.SJL-Ptrca/BoyAiTac mice (CD45.1 mice) were euthanized and spleens were collected. After tissue dissection and red blood cell lysis, primary mouse T cells were purified using the mouse Pan T cell Isolation Kit (Miltenyi Biotec). Purified T cells were cultured in RPMI-1640 (Invitrogen) supplemented with 10% FBS (HyClone), 10 mM HEPES (Invitrogen), 2 mM l-glutamine (Invitrogen), MEM non-essential amino acids 1× (Invitrogen), 55 μM β-mercaptoethanol, 1 mM sodium pyruvate (Invitrogen), 100 IU ml−1 recombinant human IL-2 (Proleukin; Novartis) and mouse anti-CD3/28 Dynabeads (Gibco) at a bead:cell ratio of 1:2. T cells were spinoculated with retroviral supernatant collected from Phoenix-ECO cells 24 h after initial T cell activation as described44 (link),45 (link) and used for functional analysis 3–4 days later.
Mouse anti cd3 28 dynabeads
Manufactured by Thermo Fisher Scientific
Mouse anti-CD3/28 Dynabeads are uniform superparamagnetic beads coated with mouse monoclonal antibodies specific for the CD3 and CD28 molecules on human T cells. These beads can be used to activate and expand T cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Mouse pan t cell isolation kit, by Miltenyi Biotec (5 mentions)
Rpmi 1640, by Thermo Fisher Scientific (5 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (5 mentions)
L glutamine, by Thermo Fisher Scientific (5 mentions)
Hepes, by Thermo Fisher Scientific (5 mentions)
Mem non essential amino acids 1, by Thermo Fisher Scientific (4 mentions)
Recombinant human il 2 proleukin, by Novartis (3 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
Proleukin, by Novartis (1 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (1 mentions)
5 protocols using mouse anti cd3 28 dynabeads
1
Isolation and Activation of Mouse T Cells
2
Isolation and Expansion of Mouse T Cells
3
Mouse T Cell Isolation and Activation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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B6.SJL-Ptrca Pepcb/BoyJ(CD45.1 mice) were euthanized and spleens were collected. After tissue dissection and red blood cell lysis, primary mouse T cells were purified using the mouse Pan T cell Isolation Kit (Miltenyi Biotec; 130-095-130). Purified T cells were cultured in RPMI-1640 (Invitrogen; 11-875-085) supplemented with 10% FBS (Corning; 35–010-CV), 10 mM HEPES (Thermo Scientific; 15630080), 2 mM L-glutamine (Thermo Scientific; 25030164), MEM non-essential amino acids 1x (Thermo Scientific; 11140076), 55 μM β-mercaptoethanol (Thermo Scientific; 21985023), 1 mM sodium pyruvate (Thermo Scientific; 11360070), 100 IU ml−1 recombinant human IL-2 (Proleukin; Novartis) and mouse anti-CD3/28 Dynabeads (Gibco; 11452D) at a bead:cell ratio of 1:2. T cells were spinoculated with retroviral supernatant collected from Phoenix-ECO cells 24 h after initial T cell activation as described79 (link),80 (link) and used for functional analysis 3–4 days later.
4
Expansion and Transduction of Mouse T Cells
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B6.SJL-Ptrca/BoyAiTac mice (CD45.1 mice) were euthanized and spleens were collected. After tissue dissection and red blood cell lysis, primary mouse T cells were purified using the mouse Pan T cell Isolation Kit (Miltenyi Biotec). Purified T cells were cultured in RPMI-1640 (Invitrogen) supplemented with 10% FBS (HyClone), 10 mM HEPES (Invitrogen), 2 mM l -glutamine (Invitrogen), MEM non-essential amino acids 1× (Invitrogen), 55 µM β-mercaptoethanol, 1 mM sodium pyruvate (Invitrogen), 100 IU ml−1 recombinant human IL-2 (Proleukin; Novartis) and mouse anti-CD3/28 Dynabeads (Gibco) at a bead:cell ratio of 1:2. T cells were spinoculated with retroviral supernatant collected from Phoenix-ECO cells 24 h after initial T cell activation as described in refs. 44 (link),45 (link) and used for functional analysis 3–4 d later.
5
Isolation and Expansion of Mouse T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
B6.SJL-Ptrca/BoyAiTac mice (CD45.1 mice) were euthanized and spleens were harvested. Following tissue dissection and red blood lysis, primary mouse T cells were purified using the mouse Pan T cell Isolation Kit (Miltenyi Biotec). Purified T cells were cultured in RPMI-1640 (Invitrogen) supplemented with 10% fetal bovine serum (FBS; HyClone), 10 mM HEPES (Invitrogen), 2 mM L-glutamine (Invitrogen), MEM nonessential amino acids 1× (Invitrogen), 55 uM β-mercaptoethanol, 1 mM sodium pyruvate (Invitrogen), 100 IU/ mL of recombinant human IL-2 (Proleukin; Novartis) and mouse anti-CD3/28 Dynabeads (Gibco) at a bead:cell ratio of 1:2. T cells were spinoculated with retroviral supernatant collected from Phoenix-ECO cells 24 hours after initial T cell expansion as described33 (link),53 (link) and used for functional analysis 3–4 days later.
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