DC-DAI-RNAi and DC-GFP were stimulated with LPS for 48 h, incubated with mitomycin C (Abmole, US) for 2 h at 37°C, and washed three times with PBS. The pretreated DCs were co-cultured with T cells from C57BL/6 mice spleen in 96-well culture plates at ratios of 1:1, 1:10, 1:20, 1:50, and 1:100 for 72 h. Untreated T cells and RPMI-1640 with 10% FBS culture medium served as negative and blank controls, respectively. Cell Counting Kit (CCK-8, Abmole, US) was added to the medium for 4 h before the end of culture, and T cell proliferation was analyzed by determining the OD450 using a microplate reader (Bio-Rad, iMark™). The stimulation index (SI) was calculated according to the following formula: stimulation index = (ODsample − ODblank control)/(ODnegative control − ODblank control). Meanwhile, MLR culture supernatants were collected from each group for concentration measurements of IL-10, IL-17, and IFN-γ using ELISA kits according to the manufacturer’s instructions.
Cell counting kit cck 8
Manufactured by AbMole
Sourced in United States
The Cell Counting Kit (CCK-8) is a colorimetric assay used to determine the number of viable cells in a sample. It utilizes a water-soluble tetrazolium salt that is reduced by dehydrogenase enzymes in living cells, producing a water-soluble formazan dye that can be measured spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Mitomycin c, by AbMole (1 mentions)
Pdms sylgard 184, by Dow (1 mentions)
Phosphate buffer, by Merck Group (1 mentions)
Fluorescein isothiocyanate (fitc), by Merck Group (1 mentions)
Acetic acid, by Sinopharm (1 mentions)
Streptozotocin (stz), by AbMole (1 mentions)
Dibasic sodium phosphate na2hpo4, by Sinopharm (1 mentions)
Genipin, by J&K Scientific (1 mentions)
2 protocols using cell counting kit cck 8
1
Modulation of T Cell Proliferation by LPS-Stimulated DCs
2
Hydrogel-based Biomaterial Characterization
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PDMS (Sylgard 184) was obtained from Dow Corning (Midland, USA). PLA was purchased from Lakeshore Biomaterials Inc. (AL, USA). Cell counting kit (CCK-8) and STZ were purchased from AbMole BioScience (TX, USA). Gel (from cold water fish skin and from porcine skin), FITC, phosphate buffer, and deuterium oxide (D2O, D > 99.9%) were purchased from Sigma-Aldrich (MO, USA). Ehrlich’s reagent was purchased from BioTechnology Co Ltd. (Shanghai, China). Acetic acid, sodium carbonate anhydrous (Na2CO3), and dibasic sodium phosphate (Na2HPO4) were obtained from Sinopharm Chemical Reagent Co Ltd. (Beijing, China). Insulin (short-, intermediate-, and long-acting types) was purchased from Wanbang Biopharmaceuticals Co. Ltd. (Jiangsu, China). HA (Mw ≈ 350,000 Da) was purchased from Toronto Research Chemicals Inc. (Toronto, Canada). BDDE (Mw = 202.25 Da) was purchased from Adamas Reagent Co. Ltd. (Basel, Switzerland). Genipin was purchased from J&K Scientific (Beijing, China). Mouse fibroblast cell lines (L-929 cells) were purchased from iCell Bioscience (Shanghai, China), and female Balb/c mice were obtained from the Institute of Laboratory Animal Sciences (Beijing, China). Aqueous solutions were carried out using ultrapure water from a reverse osmosis membrane system, unless otherwise stated.
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