Clonogenic assay was used to determine cell survival after treatment with NMDi or XR-2 alone or in combination. 22Rv1 or HCT116 cells were plated in a six-well plate at a density of 800 or 1500 cells per well, respectively. Cells were treated with chemicals at the indicated concentration 24 h after plating. The media of the cells was replaced with fresh media 24 h post-treatment, and the cells were cultured for 2 weeks. The colonies were fixed with methanol, stained with 0.2% crystal violet (Meilunbio, Dalian, China) and washed with water; subsequently, the colony images were captured.
Crystal violet
Manufactured by Meilun
Sourced in China
Crystal violet is a synthetic dye commonly used in laboratory settings. It is a dark purple crystalline solid with the chemical formula C25H30ClN3. Crystal violet is primarily used as a staining agent in microscopy and other laboratory techniques to enhance the visibility of cellular structures and microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Paraformaldehyde (pfa), by Solarbio (1 mentions)
5 ethynyl 2 deoxyuridine (edu), by Beyotime (1 mentions)
Synergy h1m multimode microplate reader, by Agilent Technologies (1 mentions)
Cck 8 reagent, by Meilun (1 mentions)
Upper chamber, by Jet Biofil (1 mentions)
96 well plates, by Wuhan Servicebio Technology (1 mentions)
Cell counting kit 8 (cck8), by Biosharp (1 mentions)
Annexin 5 fitc pi apoptosis kit, by Absin (1 mentions)
Prism 8, by GraphPad (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Meilun (1 mentions)
Ckx3 slp, by Olympus (1 mentions)
16 protocols using crystal violet
1
Clonogenic Assay for Cell Survival Analysis
2
Cell Proliferation and Colony Formation Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell proliferation assays were performed using Cell Counting Kit-8 (CCK-8, MCE, HY-K0301, Monmouth, NJ, USA). Briefly, a CCK-8 reagent was added to the culture medium according to the instructions, then placed in an incubator at 37 °C with 5% CO2 in the dark for 2 h. Finally, the absorbance at 450 nm was measured using a microplate reader.
U87 cells (1000 per well), ln229 cells (2000 per well), and U251 cells (2000 per well) were seeded in 6-cm dishes and then cultured in an incubator at 37 °C with 5% CO2. A fresh complete medium was replaced every 3 days. After 14 days of culture, cells were fixed with 4% paraformaldehyde (Solarbio, p1110, Beijing, China) and stained with 2.5% crystal violet (Meilunbio, MA0148, Dalian, China). Pictures were finally taken with a digital camera by a researcher who was blinded to the group allocation to calculate the colony formation rate. Colony formation rate = amount of colonies/number of seeded cells.
U87 cells (1000 per well), ln229 cells (2000 per well), and U251 cells (2000 per well) were seeded in 6-cm dishes and then cultured in an incubator at 37 °C with 5% CO2. A fresh complete medium was replaced every 3 days. After 14 days of culture, cells were fixed with 4% paraformaldehyde (Solarbio, p1110, Beijing, China) and stained with 2.5% crystal violet (Meilunbio, MA0148, Dalian, China). Pictures were finally taken with a digital camera by a researcher who was blinded to the group allocation to calculate the colony formation rate. Colony formation rate = amount of colonies/number of seeded cells.
3
Cell Proliferation Assays for MGC803 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The phenotype assays of MGC803 cells were performed after transfected with eccDNA for 48 h. For CCK8 testing, approximately transfected 2 × 103 cells in 100 μl were incubated in quadruplicate in 96-well plates. At 0, 1, 2, 3 and 4 day, the CCK-8 reagent (Meilunbio) was added to each well and incubated at 37 °C for 2 h. Absorbance at 450 nm was recorded by the Synergy H1M Multimode microplate reader (BioTek).
For formation analysis, approximately transfected 1 × 103 cells in 3 ml were incubated in quadruplicate in 6-well plates. Cells were stained by crystal violet (Meilunbio) followed by incubating for two weeks. Image J software (version 1.8.0; National Institutes of Health) was used for calculating the cell cluster each well.
For EdU assay, approximately transfected 8 × 103 cells in 100 μl were incubated in quadruplicate in 96-well plates. Transfected cells were added with 50 mM EdU (Beyotime) and incubated for another 2 h following by incubation for 24 h at 37 °C and 5% CO2. Cells were then fixed with 4% paraformaldehyde, permeated with 0.5 trition and stained with Apollo Dye Solution for proliferating cells. DAPI was used for labeling the Nucleic acids for all cells. Three randomly selected fields were taken using a fluorescence microscope. The cell proliferation rate was calculated using Image j software.
For formation analysis, approximately transfected 1 × 103 cells in 3 ml were incubated in quadruplicate in 6-well plates. Cells were stained by crystal violet (Meilunbio) followed by incubating for two weeks. Image J software (version 1.8.0; National Institutes of Health) was used for calculating the cell cluster each well.
For EdU assay, approximately transfected 8 × 103 cells in 100 μl were incubated in quadruplicate in 96-well plates. Transfected cells were added with 50 mM EdU (Beyotime) and incubated for another 2 h following by incubation for 24 h at 37 °C and 5% CO2. Cells were then fixed with 4% paraformaldehyde, permeated with 0.5 trition and stained with Apollo Dye Solution for proliferating cells. DAPI was used for labeling the Nucleic acids for all cells. Three randomly selected fields were taken using a fluorescence microscope. The cell proliferation rate was calculated using Image j software.
4
Cell Migration Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Migration assays were performed using 24-well transwell inserts with 8.0 μm microporous membranes (Corning, Beijing, China). THP-1 cells were preincubated with PMA (100 ng/mL) for 24 h before stimulation with LPS (100 ng/mL) for 6 h. RAW264.7 cells were treated with LPS (100 ng/mL) for 6 h. These two cell lines were starved in FBS-free DMEM for 24 h, and then treated with DPI (10-13 or 10-14 M) or DMSO for 24 h. Supernatant was collected and added to the lower chamber to serve as a chemoattractant. THP-1 and RAW264.7 cells were resuspended in 100 μL serum-free medium and then seeded into the upper chamber at a density of 4×105/well. Cells were allowed to migrate for 12 h at 37 °C in 5% CO2 and then fixed and stained with crystal violet (#MA0150, Meilunbio). The number of migrated cells in three random fields per well was counted. Images were acquired using a microscope (ZEISS, Jena, Germany) with a charge-coupled device (CCD) camera.
5
Colony Formation Assay for Transfected HCC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For colony formation, logarithmic growth phase transfected HCC cells were collected and trypsinized into single cells. In six-well plates, 1000 cells were inoculated per well. After two weeks incubation at 37 °C in 5% CO2, cell clones that had formed from individual cells were directly observed by eye, then culture medium was removed and the remaining cells were washed three times with PBS. The cells were fixed with 4% paraformaldehyde for 30 min and stained with 0.1% crystal violet (Meilunbio, Dalian, China) for 20 min at room temperature. Following staining, dishes were placed on a transparent grid and the number of clone cells was counted using a inverted microscope (Nikon ECLIPSE, Shanghai, China).
6
Cell Colony Formation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transfected MHCC97H cells in the logarithmic growth phase were seeded in six-well plates at a density of 500–1,000 cells per well. When individual cells grew into colonies that were visible by eye, cells were stained with 0.1% crystal violet (Meilunbio, Dalian, China). The number of clones was counted using an inverted microscope.
7
Microtubule Stabilization Regulates Endothelial and Pericyte Migration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The microtubule stabilization effect on migration of endothelial cells and pericytes was evaluated using transwell chambers with an 8-μm pore-sized polycarbonate filter membrane. In the Epo B group, the cells were incubated with Epo B (10 ρM, 100 ρM) for 24 h. Then cells were trypsinized and suspended in serum-free medium. The cells (1 × 105) were seeded on the upper side of chamber with the serum-free medium containing Epo B (10 ρM, 100 ρM). The lower well was filled with normal medium containing Epo B (10 ρM, 100 ρM). After being incubated for 24 h, the cells on the upper side of the membrane were gently removed. Then the cells that had migrated through the 8 μm pores were fixed with 4% paraformaldehyde and stained with crystal violet (Dalian Meilun Biology Technology Co., Ltd., China). The migrated cells were counted in five different fields under microscope and quantified by image J software (NIH, Bethesda, MD, USA).
8
Isolation and Culture of Myeloid Cells from Tumor-Bearing Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tumor-bearing mice were sacrificed, and peripheral blood samples were collected in an anti-coagulation tube. RBCs were removed by an RBC lysis buffer (Cat. No. MA0207, Meilunbio, China). Blood cells were then washed with PBS. The cell suspension was seeded onto six-well plates for 24 h, and the non-adherent cells were removed by changing the medium. Cells were cultured with 10% FBS-DMEM (Cat. No. 10099-141, Gibco; Cat. No. MA0213, Meilunbio, China) for 10 days, and stained with crystal violet (Cat. No. MA0149, Meilunbio, China) for further analysis.
9
Colony Formation Assay for HCC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transected HCC cells (1 × 103 cells each well) were added into a 6-well plate and cultured in the incubator. After two weeks incubation, the formed cell colonies were fixed with 4% paraformaldehyde for 30 min and subsequently stained with 0.5% crystal violet (meilunbio, Dalian, China) for 30 min at room temperature.
10
Cell Proliferation, Migration, and Clonogenicity Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells (8000/well) were seeded into 96-well plates (Servicebio, WuHan, China), and proliferation was evaluated using Cell Counting Kit-8 (Biosharp Biotechnology, Beijing, China) reagent. The optical density was read at 450 nm. For clone formation assays, cells were cultured for 14 days, fixed with 4% paraformaldehyde for 15 min, and stained with 0.1% crystal violet for 15 min (Meilunbio, Dalian, China). For scratch assays a linear wound was created in a monolayer of serum-starved cells using a 10-μL pipette tip, and cell coverage across this line was determined. For Transwell migration assays, cells were seeded into the upper chamber (8 μm; BIOFIL, Guangzhou, China) and incubated with serum-free medium. The lower chamber contained medium with 10% FBS. After 24 hours, cells were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet.
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!