Human peripheral blood mononuclear cells (PBMC) were isolated from the blood of adult donors obtained from the Blood Transfusion Centre (University Medical Center Freiburg). Venous blood was centrifuged on a LymphoPrep™ gradient (density: 1.077 g/cm3, 20 min, 500g, 20 °C; Progen). Afterwards cells were washed three times with PBS and cell viability and concentration was determined using the trypan blue exclusion test. Cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Invitrogen) supplemented with 10% heat-inactivated fetal bovine serum (PAA), 2 mM L-glutamine, 100 U/mL penicillin and 100 U/mL streptomycin (Invitrogen) at 37 °C in a humidified incubator with a 5% CO2/95% air atmosphere. PBMC were additionally stimulated with anti-human CD3 (clone HIT3) and anti-human CD28 (clone 28.6) mAb (100 ng/mL; both from eBioscience). Incubation was carried out as indicated in the figure captions in the presence of medium alone, different control substances, C. ipecacuanha fractions, or isolated cyclotides.
Anti human cd28 clone 28.6 mab
Manufactured by Thermo Fisher Scientific
The Anti-human CD28 (clone 28.6) mAb is a monoclonal antibody that binds to the CD28 receptor on human cells. The CD28 receptor is a co-stimulatory molecule that plays a crucial role in the activation and proliferation of T cells. This antibody can be used in various immunological applications such as flow cytometry, immunoprecipitation, and cell stimulation experiments.
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2 protocols using anti human cd28 clone 28.6 mab
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Isolation and Stimulation of PBMCs
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Isolation and Stimulation of PBMCs
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Human peripheral blood mononuclear cells (PBMC) were isolated from the blood of adult donors obtained from the Blood Transfusion Centre (University Medical Center Freiburg). Venous blood was centrifuged on a LymphoPrep™ gradient (density: 1.077 g/cm3, 20 min, 500g, 20 °C; Progen). Afterwards cells were washed three times with PBS and cell viability and concentration was determined using the trypan blue exclusion test. Cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Invitrogen) supplemented with 10% heat-inactivated fetal bovine serum (PAA), 2 mM L-glutamine, 100 U/mL penicillin and 100 U/mL streptomycin (Invitrogen) at 37 °C in a humidified incubator with a 5% CO2/95% air atmosphere. PBMC were additionally stimulated with anti-human CD3 (clone HIT3) and anti-human CD28 (clone 28.6) mAb (100 ng/mL; both from eBioscience). Incubation was carried out as indicated in the figure captions in the presence of medium alone, different control substances, C. ipecacuanha fractions, or isolated cyclotides.
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