Cells were collected in the logarithmic growth phase, and a suspension prepared with a concentration of 2000 cells/6‐well plate. The culture dish was placed in an incubator at 37°C and 5% carbon dioxide. After culturing in complete medium for 7 days, the cells were fixed with 4% paraformaldehyde for 30 min, and then stained with 0.1% crystal violet solution (Sigma‐Aldrich). After washing the excess dye with phosphate buffered saline (PBS), a microscope was used to detect the total number of colonies with a minimum of 80 cells, and the data recorded and analyzed.
Crystal violet solution
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, China, Japan, Sao Tome and Principe, Belgium, Macao, Switzerland, Australia, Israel
Crystal violet solution is a widely used laboratory reagent that serves as a staining agent. It is a purple-colored dye that has a variety of applications in various scientific fields, such as microbiology and cell biology. The solution contains a concentrated form of the crystal violet dye, which can be diluted as needed for specific experimental purposes.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by BD (41 mentions)
Paraformaldehyde (pfa), by Merck Group (24 mentions)
Acetic acid, by Merck Group (18 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (16 mentions)
Transwell chamber, by Corning (15 mentions)
Methanol, by Merck Group (13 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (11 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (11 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (11 mentions)
Formalin, by Merck Group (8 mentions)
Streptomycin, by Thermo Fisher Scientific (8 mentions)
Phosphate buffered saline (pbs), by Merck Group (8 mentions)
Multiskan fc, by Thermo Fisher Scientific (8 mentions)
Crystal violet, by Merck Group (7 mentions)
Formaldehyde solution, by Merck Group (7 mentions)
Formalin, by Fujifilm (7 mentions)
Penicillin, by Thermo Fisher Scientific (7 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (7 mentions)
Inverted microscope, by Olympus (6 mentions)
Matrigel, by Corning (6 mentions)
775 protocols using crystal violet solution
1
Quantification of Cell Colony Formation
2
Static Biofilm Quantification Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Static biofilm assays were performed as described previously (83 (link)). Bacteria were grown overnight in LB at 37°C under static conditions. Bacteria were normalized to an OD600 of 0.1 in LB broth and seeded into round bottom 96-well plates (Corning). Then, 0.2% l- arabinose was added for induction of complementation plasmids. Plates were incubated for 23 h at 37°C in static conditions and then washed (2×) with sterile water and dried for 25 min. Biofilms were fixed with methanol, stained with a 0.1% (wt/vol) crystal violet solution (Sigma), and washed (2×) with sterile water. The bound dye was solubilized with 30% acetic acid and the OD585 was measured. Each experiment was performed at least three times in technical triplicate.
3
Clonogenic Survival Assay for DNA Damage
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa cells were transduced with five different lentiviral vectors, scrambled shRNA, 53BP1 shRNA (TRCN0000018866, Sigma), Cas9-gRNA, Cas9-DN1S-gRNA, dCas9-DN1S-gRNA and DN1S, or treated with 10 µM NU7441 (Selleckchem) for 20 h. Cells transduced with empty lentiviral vectors were used as controls. Cells transduced with 53BP1 shRNA were selected in puromycin (final concentration: 5 µg/ml). Cells transduced with Cas9-gRNA, Cas9-DN1S-gRNA, dCas9-DN1S-gRNA and DN1S lentivirus vectors, all encoding the mCherry fluorochrome, were sorted for mCherry positive cells. NU7441 treated cells, puromycin selected cells, and mCherry sorted transduced cells were subjected to gamma irradiation (IR) at different doses: 0.25 Gy, 0.5 Gy, 1 Gy, 2.5 Gy and 5 Gy. IR was delivered with a J L Shepherd Mark I Model 68 A Cesium 137 irradiator. 100–200 treated cells were plated in 6 well plates, and allowed to grow. Cells that retained viability despite IR formed colonies, and after 10–14 days, colonies were stained with Crystal Violet Solution (Sigma) following the manufacturer’s instructions and counted.
4
Evaluating hGF-CM's Impact on DPSC Migration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect the effect of hGF-CM on the migration ability of DPSCs, 4 × 104 DPSCs in 100 μL serum-free αMEM were plated in the upper chamber of a Transwell with 8-μm pores (Corning), and the lower chamber (24-well plate) was injected with 350 μL either hGF-CM, 50% hGF-CM or SFM. After incubating for 6 h at 37 °C and 5% CO2, the Transwells were fixed with 4% paraformaldehyde (Merck) for 15 min and stained with 2.5 mg/mL crystal violet solution (Sigma-Aldrich, St Louis, MO, USA) in 20% methanol for 30 min. After washing the cells with PBS, the upper surface of the Transwell was cleaned with cotton swabs. Images of stained cells were taken with an inverted microscope (IX71, Olympus, Tokyo, Japan). In addition, DPSCs were stained with DAPI, and the number of migrated cells was counted by ImageJ (version 1.53t).
5
Cell Proliferation and Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A cell counting kit (CCK) for WST assay was purchased from Donginbiotech Co. (Seoul, Korea). Crystal violet solution was purchased from Sigma-Aldrich (St Louis, MO, USA). Anti-phospho-AMPK, phospho-p38, phospho-ERK, phospho-JNK, cyclin D1, CDK4, AMPK, PARP, caspase-3, Bcl-2, Bcl-xL, and Bax antibodies were purchased from Cell Signaling (Danvers, MA, USA). Anti-p38, ERK, JNK, GAPDH, and α-tubulin antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
6
Adhesion Assay for CT26 and HCT116 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To conduct the adhesion assay, CT26 and HCT116 cells (3 × 103 cells) were seeded in pre-coated Matrigel 96-well plates and treated with RA for 24 or 48 h. After incubation, the adherent cells were fixed in 10% formaldehyde for 15 min and stained with 0.05% crystal violet solution for 10 min (Sigma Chemicals, St. Louis, MO, United States). After staining, the crystal violet solution was removed and the plate was washed by PBS. After the plate was completely dried, adherent cells were observed and counted by using a microscope (Leica, Wetzlar, Germany).
7
Soft Agar Colony Formation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 5 × 103 of HME1, HME1 (AK + I), and Hela cells were mixed with 0.6% agarose (mixed with the adequate medium v:v) and were plated over a 1% agarose layer (mixed with the adequate medium v:v) (Sigma-Aldrich, St. Louis, MO, USA). The medium was renewed twice weekly by adding 200 μL of media alone to the HME1 and Hela cells and media with the adipokine and inflammatory mediators for HME1 (AK + I). After 21–30 days, colonies had formed; they were fixed using 4% PFA, washed, and further stained with 0.05% crystal violet solution (Sigma-Aldrich, St. Louis, MO, USA). The colonies were photographed at 40× magnification using a camera fitted with an inverted microscope. Cell colonies were counted manually in four randomly selected magnification fields; the data represent the number of colonies. Three independent experiments were performed.
8
Matrigel-based Invasion Assay for CRC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Invasion assays were conducted in triplicate using BioCoat™ Matrigel© invasion chambers (BD Biosciences) with pre-coated 8.0-μM mesh membrane inserts. FBS (10%) in McCoy's 5A medium was used as the chemoattractant in the outer chamber of the well for the HT-29 and HCT116 cell lines, whereas DMEM supplemented with 10% FBS was used for the three AA CRC cell lines. For all cell lines, 5×104 cells/ml were suspended in their respective medium with 0.5% FBS, seeded into the inner chamber of the well and incubated at 37°C in 5.0% CO2 for 24 h. After incubation, non-invading cells were aspirated from the chamber and the Matrigel© layer was removed from the mesh membranes. Invading cells entrapped in the membrane were fixed with 100% methanol and stained with 0.5% crystal violet solution (Sigma-Aldrich; Merck KGaA). For each membrane, 8 fields of view at ×20 under a phase-contrast microscope were quantified and averaged.
9
Transwell Invasion Assay for Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transwell assays were performed as previously described [25 (link)]. Upper chambers of Transwell inserts with 8.0 μm pore provided by Corning (Corning, NY, USA) were precoated with Matrigel (BD, Franklin Lakes, NJ, USA). HGC-27 and AGS cells were seeded on the upper chambers and cultured for 24 hours. Cells invaded into the lower chambers, which were fixed and stained with 1% crystal violet solution (Sigma, St. Louis, MO, USA) for imaging under a microscope from Olympus.
10
Colony Formation Assay with Vemurafenib and Ascorbate
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the colony formation assay, 1.0 × 103 cells were seeded into cavities of a 12-well plate and incubated overnight. Subsequently, the cells were treated with indicated concentrations and combinations of vemurafenib and ascorbate for 7 days. Treatment was carried out in triplicates and DMSO (0.02%) was used as a solvent control. To analyze the number of formed colonies, the cells were fixed in 4% formalin and stained with 3% crystal violet solution (Sigma-Aldrich; Taufkirchen, Germany) in 80% methanol for 2 h.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!