The effect of IFN-λ1 to keratinocyte proliferation was measured by Cell Counting kit (CCK-8) assay. Keratinocytes were seeded in a 96-well plate (about 3 × 103 cells per well). After 24 h incubation, IFN-λ1 at the concentrations of 1 ng/ml, 10 ng/ml and 100 ng/ml (Pereprotech, AF-300-02L, United States) were added to the culture media. 10 μl of TransDetect® Cell Counting Kit (CCK) (Transgen, FC101-01, China) was added to each well at the time of 0, 24, 48, and 72 h, respectively. After 1 h incubation, the absorbance at 450 nm was measured by the Multiskan™ FC Microplate Photometer (Thermo Scientific, 51119180, Belgium). Cell colony formation assay was also performed to verify the effect of IFN-λ1 on keratinocyte proliferation. Keratinocytes were planted to 6-well plate at 2,000 cells each well and cultured with various concentration of IFN-λ1 for 15 days. Phosphate buffered saline containing 5% trehalose (5% trehalose-PBS) (MultiSciences, 79-PD0021, China) was used as negative control. After 15 days culture, the cells were fixed with formaldehyde, stained with 0.1% crystal violet, washed with 33% glacial acetic acid and measured with a Multiskan FC microplate photometer (Thermo Scientific, 51119180, Belgium).
Transdetect cell counting kit
Manufactured by Transgene
Sourced in China
The TransDetect® Cell Counting Kit is a laboratory equipment product designed to perform accurate cell counting. It provides a reliable method to determine the number of cells in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Cell cycle analysis kit, by Thermo Fisher Scientific (2 mentions)
C10310 edu apollo in vitro imaging kit, by RiboBio (2 mentions)
Multiskan fc microplate photometer, by Thermo Fisher Scientific (1 mentions)
Cell incubator, by Sanyo (1 mentions)
Polyvinyl alcohol (pva), by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Wisent (1 mentions)
Bradford protein assay kit, by Sangon (1 mentions)
Trypsin, by Wisent (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Microtiter plate reader, by Tecan (1 mentions)
Microplate reader, by Tecan (1 mentions)
Prism v6, by GraphPad (1 mentions)
Epoch microplate spectrophotometer, by Agilent Technologies (1 mentions)
Matrigel, by BD (1 mentions)
Inverted microscope imaging system, by Olympus (1 mentions)
Three step set, by Thermo Fisher Scientific (1 mentions)
Svec4 10 cells, by Korean Cell Line Bank (1 mentions)
Spectramax plus, by Molecular Devices (1 mentions)
Spss 19, by IBM (1 mentions)
Multiskan mk3, by Thermo Fisher Scientific (1 mentions)
27 protocols using transdetect cell counting kit
1
Evaluating IFN-λ1's Impact on Keratinocyte Proliferation
2
Measuring FGSC Viability In Vitro
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The viability of FGSCs in vitro was measured using the TransDetect Cell Counting Kit (CCK) (TransGen Biotech). A cell suspension was inoculated into a 96-well plate. After adding the 10% CCK solution, the cells were incubated in the dark for 4 h in a cell incubator (Sanyo, Hong Kong, China). The absorbance of the culture plate at 450 nm was measured using a BioTek microplate reader (Bio-Rad, Hercules, CA, USA).
3
PLGA-Based Biomaterial Characterization
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PLGA (L/G = 50/50; MW = 30 kDa) was purchased from Jinan Daigang Biomaterial (Shandong, China). TF was supplied by Solarbio (Beijing, China). CMC [Viscosity: 10 mPa.s ~ 80 mpa.s (cps); MW = 200 kDa] was purchased from meilunbio (Dalian, China). TransDetect Cell Counting Kit (CCK) was obtained from TransGen Biotech Co Ltd., (Beijing, China). Dimethylsulfoxide (DMSO), N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), 2′, 7′-dichlorofluorescin diacetate (DCFH-DA) and polyvinyl alcohol (PVA) were purchased from Sigma-Aldrich (St. Louis, USA). Bradford protein assay kit was purchased from Sangon Biotech Co., Ltd (Shanghai, China). Acridine orange/ethidium bromide (AO/EB) was supplied by Aladdin (Shanghai, China). Trypsin, fetal bovine serum (FBS), and Dulbecco's minimum essential medium (DMEM) were provided by Wisent (Nanjing, China).
4
Preadipocyte Proliferation Assay
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Preadipocytes were cultured in 96-well plates at a cell density of 1 × 104 cells/well. Cell proliferation was detected after 48 h of transfection using the Trans Detect Cell Counting Kit (CCK) (TransGen Biotech Ltd., Beijing, China). Thereafter, CCK solution (10 μL) was added to wells, and the plates were incubated at 37 °C for 4 h under 5% CO2 atmosphere.
5
Splenocyte Proliferation Assay for Rift Valley Fever Virus
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In vitro lymphocyte proliferation studies were performed with splenocytes from immunized mice. Two weeks after the third immunization, three mice from each group were randomly selected and euthanized. Harvested spleens were transferred to a tissue culture dish and mechanically minced followed by treatment in red blood cell lysing agent. The resulting splenocytes were washed with RPMI 1640 medium (Gibco, United States) containing 10% FBS. Next, the splenocytes were seeded in 96-well plates to a density of 2 × 103 cells/well (100 μL/well) and stimulated for 24 h with 10 μg/mL purified RVFV Gn head protein at 37°C and 5% CO2. Lymphocyte proliferation was monitored according to the technical manual of the TransDetect Cell Counting kit (CCK) (TransGen Biotech, China).
6
Proliferation Rate of Induced Adipose-Derived Stem Cells
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The proliferation rate of IPCs was determined by TransDetect cell counting kit (CCK) (TransGen Biotech) according to the manufacturer’s instruction. On the 10th day after the induction of ADSCs, each group of cells was digested and plated at 5000 cells per well of a 96-well plate, and each group of cells was provided with 4 replicate wells and 1 zero-adjusted well (zero-well plus CCK plus medium). PBS was added to the peripheral wells for containing cell wells and placed in a layer of the incubator. The optical density at 450 nm (OD450) was measured by SpectraMaxParadigm enzyme labelling apparatus (Molecular Devices LLC, Sunnyvale, CA, USA) at 24, 48, and 72hrs, respectively. One hour before the measurement, the cells were washed with PBS, and 10 μL of CCK reagent and 90 μL of the medium were added to each well and then incubated in an incubator for 1 hr. No bubbles were observed in each well during the measurement. The final CCK value is the measured value minus the value of the zeroing well.
7
Oridonin's Cytotoxic Effect on HepG2 Cells
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The viability of cells was measured with TransDetect™ Cell Counting Kit (CCK, catalog no. FC101-01; Beijing Transgen Biotech Co., Ltd., Beijing, China) according to the manufacturer's protocol (Beijing Transgen Biotech Co., Ltd.) with minor modifications. Briefly, 100 µl HepG2-mtGrx1-roGFP2 cell suspension was dispensed into 96-well plate (5×103 cells/well) and the plate was pre-incubated at 37°C for 24 h, followed by treatment with various concentrations of oridonin (0, 5, 10, 20, 30 or 60 µM) for 8 h. Next, 10 µl (100 µl/ml) of CCK working solution was added to each well of the plate and then incubated at 37°C for 2 h. The absorbance of each well at 450 nm was measured under a multimode plate reader (PerkinElmer, Inc., Waltham, MA, USA). The results representing the average of 5 parallel samples were expressed as the relative percentage of cell growth inhibition.
8
Lymphocyte Proliferation Assay for RVFV
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In Vitro lymphocyte proliferation studies used splenocytes from immunized mice. The spleen was treated as in the ELISpot experiment. Splenocyte cultures were seeded in 96-well plates at 100 μL per well (2 × 103 cells) and then stimulated with purified RVFV eGn protein (10 μg/mL) at 37 °C in 5% CO2 for 24 h. Lymphocyte proliferation was monitored according to the instructions in the technical manual of the commercial reagent TransDetect Cell Counting kit (CCK) (Transgen biotech, Beijing, China). The CCK solution (10 µL) was directly added to each well, the incubation was continued for 3 h, and then the absorbance was measured at 450 nm.
9
Cytotoxicity Evaluation of Quercetin
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Trans Detect® Cell Counting Kit (CCK, TransGen Biotech, Beijing, China) was used for cytotoxicity assay. The viability of RD cells treated with different concentrations of quercetin was determined. Briefly, the RD cells were inoculated into the cell culture plate 1 day in advance, and the medium was discarded the next day, followed by the addition of quercetin at different concentrations. After 48 h of treatment, 10 μL CCK solution was added to each well, cultured for 1–4 h, and the absorbance value was measured at 450 nm.
10
Dose-Dependent Antiviral Activity Assay
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Dose response experiments were performed to test CC50s and EC50s of drugs as described above with minor modifications. For CC50 assays, 96 μL of F81 cells (2.5 × 104 cells/well) was mixed with 4 μL prediluted drugs at final concentrations ranging from 0.0024–160 μM. For EC50 assays, 86 μL of F81 cells (2.5 × 104 cells/well) was pretreated with 4 μL prediluted drugs at final concentrations ranging from 0.0024–160 μM for 1 h, and then treated cells were infected with 10 μL CPV at an MOI of 0.076. Both CC50 and EC50 assays were done in triplicate. Cell cytotoxicity and inhibition of CPV infection were both examined after 40 h incubation using the TransDetect® Cell Counting Kit (TransGen Biotech, Beijing, China). The CC50 and EC50 values were calculated via a best-fit Log (dose)-response curve-fitting in GraphPad Prism software (version 7.00, La Jolla, CA, USA) [21 (link)].
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