Cells were seeded at 1×105 cell/well in 24-well plates. A Cell Counting Kit 8 (CCK-8) assay was used to measure the proliferation of cells in each group. The CCK-8 solution from the Cell Counting Kit (Beijing Solarbio Science & Technology, Co., Ltd., Beijing, China) was added to each well (10 µl/well). Cells were then maintained at 37°C for 4 h. The absorbance of the reaction regent was determined at 450 nm using a microplate reader (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
Cell counting kit
Manufactured by Solarbio
Sourced in China
The Cell Counting Kit is a lab equipment designed to quantify the number of cells in a sample. It provides a standardized method for determining the concentration of cells in a suspension.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Bio-Rad (1 mentions)
Ab237526, by Abcam (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Recombinant rnase inhibitor, by Takara Bio (1 mentions)
Sybr fast qpcr master mix, by Roche (1 mentions)
Goat anti mouse igg, by OriGene (1 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions)
Primescript 2 rtase, by Takara Bio (1 mentions)
Trypsin, by Solarbio (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Transwell chamber, by Corning (1 mentions)
Enzyme labeling instrument, by Thermo Fisher Scientific (1 mentions)
5 protocols using cell counting kit
1
Cell Proliferation Assay using CCK-8
2
TGF-β1 Concentration-Dependent Cell Proliferation
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MDA-MB-231 and MCF-7 cells were cultivated in 96-well plates and under the conditions of 37°C and 5% CO2. On the second day, different concentrations of TGF-β1 protein (5, 10ng/ml) were added for intervention. Finally, the cell counting kit (Solarbio, Beijing, China) was used to measure the absorbance of OD450 at different times (24, 48 and 72h). CCK-8 (10μL) was added to each well and we should pay attention to operate in the dark during the whole process. After incubating at 37°C for 2 h, the absorbance at 450 nm was detected with a microplate reader.
3
VSMC Proliferation Assay
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VSMC proliferation was analyzed by using a cell counting kit (Solarbio, Beijing, China) according to the manufacturer's instruction. In brief, VSMCs were incubated in 96-well plates (0.5 × 104 cells per well) with physiological saline, GSK, PDGF-BB or PDGF-BB with GSK and then cultured for 48 h. Ten microliters of CCK-8 agent was then added to each well and incubated with VSMCs for another 2 h at 37 °C. By using a microplate reader, VSMC proliferation was finally determined via calculating the relative absorbance at 450 nm.
4
Osteosarcoma Cell Line Characterization
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The osteosarcoma cell line MG63 was sourced from the Shanghai Cell Bank, Chinese Academy of Sciences. PRR11 rabbit anti-human polyclonal antibody (Ab237526) was bought from the Abcam Company, USA. Trizol was bought from Ambion, USA. Reverse transcription kit PrimeScript II RTase and Recombinant RNase Inhibitor were bought from TAKARA. Goat anti-mouse IgG, goat anti-rabbit, and DAB chromogenic kits were bought from OriGene, USA. SYBR FAST qPCR Master Mix was bought from KAPABiosystems. Lipofectamine 2000 was bought from Invitrogen, USA. MEM, Opti-MEM, and fetal bovine serum were bought from Gibco, USA. PBS, 0.25% trypsin, and the cell counting kit were bought from Beijing solarbio. Transwell chambers were bought from Corning, USA.
5
Cell Viability Assay of Adipose Fibroblasts
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Cell counting kit (Solarbio) was utilized as described by the manufacturer's instruction. Briefly, AFs were cultured in 96-well plastic plates in complete media at an initial density of 5 × 10 4 /mL. After treatment of SKL2001 or insulin, 10 uL of CCK-8 reagent was added to each well and incubated with AFs at 37°C for another 2 h. Finally, the viability of AFs was determined by Enzyme Labeling Instrument (Thermo Fisher Scientific, Waltham, MA, USA) via calculating the relative absorbance at 450 nm.
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