A549 and H1299 cells were stably transfected BLK and control. The cells were planted into a 96-well plate added with 100 µl DMEM containing 10% FBS. Cell Counting Kit (CCK)-8 reagent (Beyotime Biotech) was used to analyze the cell proliferation experiment. Cell proliferation was observed at 0h, 24h, 48h, 72h. Briefly, each well was filled with 10µl CCK-8 solution, and the plate was incubated at 37°C for 2 h. Multiscan MK3 microplate reader (Themo Fisher Scientific, Inc.) was used to determine the absorbance of the cells at 450 nm.
Multiscan mk3 microplate reader
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Multiscan MK3 microplate reader is a compact and versatile instrument designed for a wide range of microplate-based assays. It provides accurate absorbance measurements across a broad wavelength range, enabling researchers to perform diverse applications in life science laboratories.
Automatically generated - may contain errors
Lab products found in correlation
Cell counting kit 8 reagent, by Beyotime (4 mentions)
Elisa microplate, by Corning (2 mentions)
Facscanto 2 cytometer, by BD (2 mentions)
Celltiter 96 aqueous mts reagent, by Promega (2 mentions)
Ckx41 inverted optical microscope, by Olympus (2 mentions)
Electrophoresis instrument, by Bio-Rad (2 mentions)
Cck 8, by GLPBIO (2 mentions)
7 protocols using multiscan mk3 microplate reader
1
Cell Proliferation Assay for A549 and H1299 Cells
2
Cell Proliferation Assay with CCK-8
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To examine the cell proliferation experiment, Beyotime Biotech's Cell Counting Kit (CCK)-8 reagent (Shanghai, China) was used. Proliferation of cells was seen at 0, 24, 48, and 72 h. In summary, following the addition of 10 µl of CCK-8 solution to each well, the plate was incubated at 37°C for 2 h. The cells' absorbance at 450 nm was measured using a Themo Fisher Scientific, Inc. (Waltham, MA, USA) Multiscan MK3 microplate reader.
3
Indirect ELISA for Glycosylated Samples
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In indirect ELISA, the glycosylated samples were diluted using 50 mM Na2CO3-NaHCO3 (CBS) buffer (pH 9.6) to 5 μg/mL and coated in a Corning-Costar ELISA microplate (Corning, NY, USA) with 100 μL per well at 4 °C overnight. Following this, the plates were blocked with 1% (w/v) BSA (200 μL/well) at 37 °C for 2 h. These plates were then incubated with rat antisera (diluted 200,000), rabbit antisera (diluted 320,000), and TM monoclonal antibodies (diluted 1:20,000) against TM for 1 h. In addition, polled shrimp-allergic patients’ sera diluted 10 with PBST at 37 °C for 1.5 h diluted 10-fold with PBST were added and incubated at 37 °C for 1.5 h. These plates were subsequently washed and incubated with HRP-labeled rabbit anti-rat IgG (100 μL, diluted 1:10,000), HRP-labeled goat anti-rabbit IgG (100 μL, diluted 1:10,000), HRP-labeled rabbit anti-mouse IgG (100 μL, diluted to 1:10,000) or HRP-labeled goat anti-human IgE (100 μL, diluted 1:3000) for 1 h at 37 °C. Afterward, the enzyme reaction was initiated upon adding the TMB solution and stopped after adding sulfuric acid. The absorbance was measured at 450 nm using a MultiscanMK3 microplate reader (Thermo Fisher Scientific Inc., Waltham, MA, USA).
4
BJE and PTX Cytotoxicity and Apoptosis
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In all the cell experiments, BJE was dissolved in dimethylsulfoxide (DMSO) and then diluted with completed DMEM medium. The final concentration of DMSO was no more than 0.1%. For cytotoxicity test, cells were seeded in 96-well plates at a density of 3 × 103 cells/ml (sextuple in each group), and treated with BJE (0.78 to 200 μg/ml) or PTX (7.8 to 2000 ng/ml). After 48 h, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT, 5 mg/ml) was added to each well followed by 4 h incubation, and the optical density was measured at 490 nm by a Multiscan MK3 microplate reader (Thermo Fisher, USA).
For experiments except cytotoxicity test, cells were seeded in 6-well plates at a density of 1 × 105 cells/ml (triplicate in each group), and treated with BJE (2.61, 5.21, 10.42 μg/ml) or PTX (26.04 ng/ml) for 48 h, and then the morphology of cell was captured by a light microscope (Olympus, Tokyo, Japan). For apoptosis assay, cells were harvested, washed with cold PBS, and stained with Annexin V (2.5 μL/test)/fluorescein isothiocyanate (FITC, 5 μL/test) (Lianke Biotech, Co., Ltd., 82480552) for 5 min at RT in the dark. Cell apoptosis was measured with a FACS Canto II cytometer (BD, USA), and the data was analyzed by Diva software (version 6.0).
For experiments except cytotoxicity test, cells were seeded in 6-well plates at a density of 1 × 105 cells/ml (triplicate in each group), and treated with BJE (2.61, 5.21, 10.42 μg/ml) or PTX (26.04 ng/ml) for 48 h, and then the morphology of cell was captured by a light microscope (Olympus, Tokyo, Japan). For apoptosis assay, cells were harvested, washed with cold PBS, and stained with Annexin V (2.5 μL/test)/fluorescein isothiocyanate (FITC, 5 μL/test) (Lianke Biotech, Co., Ltd., 82480552) for 5 min at RT in the dark. Cell apoptosis was measured with a FACS Canto II cytometer (BD, USA), and the data was analyzed by Diva software (version 6.0).
5
Cell Viability Assay with MTS Reagent
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The cells were inoculated in 96-well plates at a density of 3 × 104 cells/mL and treated with the corresponding drugs. After 48 h, 20 μL of Cell Titer 96®AQueous MTS Reagent (Promega, Wisconsin, United States) was added to each well, and the optical density was measured at 490 nm on a Multiscan MK3 microplate reader (Thermo Fisher, United States) after 4 h.
6
Multimodal Experimental Techniques
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The instruments used included a multiscan MK3 microplate reader (Thermo, USA), CKX-41 inverted optical microscope (Olympus, Japan), TB-718 biological tissue embedding machine (Hubei Taiwei Electronics Company), NOVA ultrathin slicer (LKB Company), electrophoresis instrument (Bio-Rad, USA), Flour Chem M Gel Imaging system (Protein Simple, USA), and Quantstudio Multiple Real-Time PCR System (Life Technologies, USA).
7
Cell Viability Assay with IPyA
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4T1 cells or EMT6 cells were seeded in 96-well plates at a density of 1 × 104 cells/mL (100 μL/well) with complete medium. After adherence overnight, cells were treated with IPyA (0 ~ 4 mM) for 48 h. Next, the medium was replaced with complete medium containing 10% CCK-8 (GlpBio Technology, Montclair, CA, USA), and the optical density at 490 nm was measured with a Multiscan MK3 microplate reader (Thermo Fisher, USA) 2 h later.
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