CD4+CD25− T cells (Teff) and CD4+CD25+ T cells (Tregs) were cocultured in 96-well plates coated with 50 ng/mL anti-CD3 mAb (eBioscience, USA) at a density of 104 cells/well with different Teff/Treg ratios (1 : 1, 1 : 1/2, 1 : 1/4, and 1 : 1/8). All wells were cultured in a final volume of 200 μL with the presence of T cell-depleted and irradiated antigen presenting cells (105 cells/well). After 72 h, [3H]-thymidine (1 μCi/well) was added for 16 h prior to the determination of proliferation by scintillation counting (MicroBeta1450 Liquid Scintillation Counter; Perkin Elmer, USA). Percent inhibition of proliferation was determined as follows: (1-[3H]-thymidine uptake of cocultured Treg and Teff)/Teff alone × 100%. Triplicate wells were used in all suppression experiments.
Microbeta1450 liquid scintillation counter
Manufactured by PerkinElmer
Sourced in United States
The MicroBeta1450 Liquid Scintillation Counter is a versatile instrument used for the detection and quantification of radioactive samples. It utilizes liquid scintillation technology to measure the emissions from radioactive isotopes. The MicroBeta1450 is capable of handling a variety of sample types and provides accurate and reliable results.
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Lab products found in correlation
2 protocols using microbeta1450 liquid scintillation counter
1
Treg-Mediated Suppression of T-Cell Proliferation
2
Quantifying cAMP Synthesis in Receptor Cells
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EXAMPLE 9
Functional Assay-cAMP Synthesis
The ability of glucagon analogs to induce cAMP was measured in a firefly luciferase-based reporter assay. HEK293 cells co-transfected with a receptor (glucagon receptor, GLP-1 receptor or GIP receptor) and luciferase gene linked to cAMP responsive element were serum deprived by culturing 16 h in DMEM (Invitrogen, Carlsbad, Calif.) supplemented with 0.25% Bovine Growth Serum (HyClone, Logan, Utah) and then incubated with serial dilutions of either glucagon, GLP-1, GIP or novel glucagon analogs for 5 h at 37° C., 5% CO2 in 96 well poly-D-Lysine-coated “Biocoat” plates (BD Biosciences, San Jose, Calif.). At the end of the incubation 100 microliters of LucLite luminescence substrate reagent (Perkin-Elmer, Wellesley, Mass.) were added to each well. The plate was shaken briefly, incubated 10 min in the dark and light output was measured on MicroBeta-1450 liquid scintillation counter (Perkin-Elmer, Wellesley, Mass.). Effective 50% concentrations were calculated by using Origin software (OriginLab, Northampton, Mass.
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