For overexpression studies, TRIB2 was subcloned into pHAGE-CMV-IRES-ZsGreen-W (Addgene, #26532). HEK-293T cells were transfected with this vector, along with psPAX2 (Addgene, #12260) and pMD2.G (Addgene, #12259) vectors using FuGENE (Promega, E2311). 72 h after transfection, the supernatant containing lentiviral particles was collected, filtered through a 0.45-μm syringe filter (Millipore) and concentrated using PEG-it solution (System Biosciences, LV810A-1). For lentiviral transduction, human naive CD4+ or CD8+ T cells were activated with anti-CD3/anti-CD28 Dynabeads (1:2) and transduced with the concentrated lentivirus in the presence of 8 μg/mL polybrene (Sigma Aldrich, TR-1003-G) and 10 U/mL human IL-2 (PeproTech, #200–02). After 1.5 days, lentivirus-containing medium was exchanged, and cells were transferred into new wells coated with anti-CD3/anti-CD28 antibodies (5 μg/mL).
Tr 1003 g
Manufactured by Thermo Fisher Scientific
The TR-1003-G is a compact and versatile laboratory centrifuge capable of processing various sample types. It features a speed range of up to 15,000 rpm and can accommodate rotors for different tube sizes. The centrifuge provides consistent and reliable performance to support routine laboratory workflows.
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2 protocols using tr 1003 g
1
Lentiviral Overexpression of TRIB2 in T Cells
2
Lentiviral Overexpression of TRIB2 in T Cells
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For overexpression studies, TRIB2 was subcloned into pHAGE-CMV-IRES-ZsGreen-W (Addgene, #26532). HEK-293T cells were transfected with this vector, along with psPAX2 (Addgene, #12260) and pMD2.G (Addgene, #12259) vectors using FuGENE (Promega, E2311). 72 h after transfection, the supernatant containing lentiviral particles was collected, filtered through a 0.45-μm syringe filter (Millipore) and concentrated using PEG-it solution (System Biosciences, LV810A-1). For lentiviral transduction, human naive CD4+ or CD8+ T cells were activated with anti-CD3/anti-CD28 Dynabeads (1:2) and transduced with the concentrated lentivirus in the presence of 8 μg/mL polybrene (Sigma Aldrich, TR-1003-G) and 10 U/mL human IL-2 (PeproTech, #200–02). After 1.5 days, lentivirus-containing medium was exchanged, and cells were transferred into new wells coated with anti-CD3/anti-CD28 antibodies (5 μg/mL).
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