300 µl of PANC1 and ASPC1 cell cultures were seeded into the upper chamber of the transwell chamber (8 µm in size, Corning, NY, USA), and 700 µl of complete medium was added to the lower chamber. After 24 h of incubation at 37 °C in 5% CO2, the cells were fixed and stained with 4% paraformaldehyde and 0.1% crystalline violet (Sigma-Aldrich, USA). Then the number of invading cells was observed under the optical microscope.
Crystalline violet
Manufactured by Merck Group
Sourced in United States
Crystalline violet is a chemical compound that is commonly used as a laboratory dye. It is a dark purple crystalline solid with the chemical formula C₂₅H₃₀ClN₃. The compound is soluble in water and various organic solvents, and it is often used in biological and chemical applications where a purple or violet staining agent is required.
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Lab products found in correlation
5 protocols using crystalline violet
1
Transwell Invasion Assay for Cell Lines
2
Transwell Assay for Cell Migration
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Cell migratory capacity was gauged employing the transwell assay. Briefly, 2 × 104 cells were inoculated into the upper chamber after resuspension with serum-free medium, and 20% FBS medium was added to the lower chamber before incubation was initiated. After 24 h of incubation, the cells were fixed and then stained with 0.1% crystalline violet (Sigma, USA). Migrated cells were counted and imaged.
3
Evaluating NCAPG2 Knockdown in Pancreatic Cancer
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The shRNA-transfected PANC1 and MIAPaCa-2 cells were fully digested, inoculated into 96-well culture plates at 2 x 10^3 cells per well and incubated at 37°C. The absorptivity was measured every day using the CCK-8 kit, for a total of seven days of incubation. The above procedures were carried out at 450 nm on the advice of the reagent supplier and four secondary wells were retained for the experimental sessions. A total of 750 transfected control or NCAPG2 shRNA-transfected PANC1, MIAPaCa-2 cells were positioned on a fresh six-well plate and cultured for 2 weeks as described in the detail culture procedure above, during which times the medium was changed every 4 days. Formed colonies were fixed in Methanol and stained with Crystalline Violet (Sigma-Aldrich) after meeting a scheduled period, then counted manually and photographed.
4
Clonogenic Assay for PANC1 and ASPC1 Cells
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PANC1 and ASPC1 cells pretreated with different vectors were inoculated in 6-well plates, incubated in an incubator (37 °C, 5% CO2) for 10 days and fixed in methanol. Afterwards, 0.1% crystalline violet (Sigma-Aldrich, USA) staining was performed. The colonies were observed and stained under the microscope after 3 times of PBS rinsing.
5
Chromatogram Visualization Using Diverse Agents
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Processes of spot visualization were carried out using several visualization agents procured from different suppliers. The solutions of these reagents were prepared as follows:
-(i) rhodamine B (POCh), (ii) Janus blue (Michrom, UK), (iii) methyl green (Fluka, Switzerland), (iv) brilliant green (POCh), (v) crystalline violet (Sigma-Aldrich), (vi) alkaline blue (Merck), (vii) gentian violet (Fluka), and (viii) methylene violet (Michrom) were used as 0.50 mg mL -1 solutions in distilled water, -fuchsine procured from Serva (Germany) was used as 0.150 mg mL -1 solution in distilled water, -brilliant cresyl blue supplied by Michrom was used as 0.50 mg mL -1 solution in 2 % aqueous NaOH.
Taking into account the visualization manner, all obtained chromatograms have been divided into four groups. The first group was dipped in an individual visualization agent for 5 s and then left at room temperature until dry. The second group was immersed in an individual visualization agent for 5 s and then dried in a laboratory dryer at 110 °C for 1 hour. The third group was sprayed with a visualization agent and left at room temperature until dry. The fourth group was sprayed with a visualization agent and dried for one hour in a laboratory dryer at 110 °C.
-(i) rhodamine B (POCh), (ii) Janus blue (Michrom, UK), (iii) methyl green (Fluka, Switzerland), (iv) brilliant green (POCh), (v) crystalline violet (Sigma-Aldrich), (vi) alkaline blue (Merck), (vii) gentian violet (Fluka), and (viii) methylene violet (Michrom) were used as 0.50 mg mL -1 solutions in distilled water, -fuchsine procured from Serva (Germany) was used as 0.150 mg mL -1 solution in distilled water, -brilliant cresyl blue supplied by Michrom was used as 0.50 mg mL -1 solution in 2 % aqueous NaOH.
Taking into account the visualization manner, all obtained chromatograms have been divided into four groups. The first group was dipped in an individual visualization agent for 5 s and then left at room temperature until dry. The second group was immersed in an individual visualization agent for 5 s and then dried in a laboratory dryer at 110 °C for 1 hour. The third group was sprayed with a visualization agent and left at room temperature until dry. The fourth group was sprayed with a visualization agent and dried for one hour in a laboratory dryer at 110 °C.
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