Crystalline violet

Manufactured by Beyotime

Sourced in China

Crystalline violet is a chemical compound commonly used in laboratory settings. It is a synthetic dye that appears as a dark purple crystalline solid. The core function of crystalline violet is to serve as a staining agent for microscopy and diagnostic applications. It can be used to stain biological samples, such as cells or tissues, for visual identification and analysis.

Automatically generated - may contain errors

Lab products found in correlation

Matrigel, by Corning (1 mentions) 8 μm pore transwell migration chambers, by Corning (1 mentions) Edu cell proliferation assay kit, by RiboBio (1 mentions) Inverted fluorescence microscope, by Olympus (1 mentions) 24 well transwell chamber, by BD (1 mentions) Matrigel, by BD (1 mentions) Paraformaldehyde (pfa), by Beyotime (1 mentions)

6 protocols using crystalline violet

Anchorage-independent Growth Assay

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The treated MCF-7 cells (1 × 103) were sorted in the culture dish pre-coated with
0.6% agar (the medium was supplemented with 20% FBS), and cultured in a 37°C
incubator with 5% CO₂. After 14 days, the cells were stained using
0.04% crystalline violet (Beyotime Biotechnology Co., Ltd) for 40 min, and the
colonies were documented and counted under an inverted microscope (Ti2, Nikon,
Tokyo, Japan).

Wu M., Wen L., Zhou Y, & Wu W. (2022). Role of lncRNA AGAP2-AS1 in Breast Cancer Cell Resistance to Apoptosis by the Regulation of MTA1 Promoter Activity. Technology in Cancer Research & Treatment, 21, 15330338221085361.

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Transwell Assay for Cell Migration and Invasion

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For Transwell plate migration assays, a total of 8 × 10⁴ cells were seeded in the upper chamber of an 8 μm Transwell plate (BD Biosciences, USA) with 100 μl of serum-free medium. In the lower chamber, 500 μl of medium containing 15% FBS was added. After 24 and 48 h of incubation, the cells in the upper chamber were carefully removed. Cells adhering to the membrane were fixed in 4% paraformaldehyde for 15 min and stained with 0.1% crystalline violet (Beyotime, China) for 15 min. For invasion assays, 50 µl of Matrigel (Corning, USA) diluted 1:4 with DMEM/F-12 was precoated in the upper chamber and seeded with 8 × 10⁴ cells in 100 µl of DMEM/F-12. The rest of the procedure was similar to that of the Transwell migration assay.

Bai M., Jiang N., Fu W., Huang C., Tian L., Mi N., Gao L., Ma H., Lu Y., Cao J., Zhang C., Yue P., Zhang Y., Lin Y., Meng W, & Li X. (2023). Establishment and characterization of a novel hilar cholangiocarcinoma cell line, CBC3T-1. Human Cell, 37(1), 364-375.

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Exosome-Mediated Schwann Cell Proliferation and Migration

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Cell proliferation assays were performed using a 5-ethynyl-2-deoxyuridine (EdU) cell proliferation assay kit (R11053.9, RiboBio, Guangzhou, China) according to the manufacturer’s instructions. Schwann cells were cultured in 96-well plates and spiked with 40 μg/ml of hAMSCs-exo or SCLCs-exo or an equal volume of PBS. After 6 h, the cells were observed under an inverted fluorescence microscope (Olympus, Tokyo, Japan).
Cell migration experiments were performed using 8-μm-pore Transwell migration chambers (Corning, NY, USA). A total of 2 × 10⁵ Schwann cells in 200 μl of serum-free medium were inoculated in the upper chamber of the Transwell plate. Subsequently, 40 μg/ml of hAMSCs-exo or SCLCs-exo or an equal volume of PBS was added to the upper chamber, while the lower chamber was filled with complete medium. The plates were incubated for 24 h before removing non-migrated cells from the upper chamber with a cotton swab. The migrated cells were stained with 0.1% crystalline violet (Beyotime, Nantong, China) for 30 min, photographed and counted under a light microscope (Olympus, Tokyo, Japan).

Hu T., Chang S., Qi F., Zhang Z., Chen J., Jiang L., Wang D., Deng C., Nie K., Xu G, & Wei Z. (2023). Neural grafts containing exosomes derived from Schwann cell-like cells promote peripheral nerve regeneration in rats. Burns & Trauma, 11, tkad013.

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Quantifying Cell Colony Formation

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Approximately 500 cells were inoculated into 6-well plates and incubated in a constant temperature incubator at 37°C with 5% CO2 for 2 weeks. The culture medium was removed, cells were washed with PBS, 4% paraformaldehyde was added to x the cells, paraformaldehyde was discarded, and crystalline violet (Beyotime, China) staining was performed to observe the number of cell colonies.

Zhao J., Shen J., Mao L., Yang T., Liu J, & Hongbin S. (2024). Cancer associated fibroblast secreted miR-432-5p targets CHAC1 to inhibit ferroptosis and promote acquired chemoresistance in prostate cancer. Oncogene, 43(27).

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Evaluating Transwell Cell Migration

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To detect cell migration capability, transwell chambers coated with and without Matrigel (Becton Dickinson, CA) were employed and utilized a 24-well transwell chamber (8

\mu

m; BD Biosciences). After 48 h of transfection, cells were inoculated into a 24-well plate. Each well was inoculated with 8

\times

10⁴ cells. TNBC cells with different plasmids or miRNA mimics were deposited in the upper chamber in serum-free medium and 10% fetal bovine serum in the lower chamber. After 24 h, the migration cells were treated for 1 h with paraformaldehyde (4%; Beyotime) and crystalline violet (Beyotime), followed by taking pictures and counting with a microscope (100 cells) from five areas randomly selected.

Jiang X., Lin J, & Zhu Z. (2024). Long-chain noncoding RNA LINC01569 upregulates filamin A-interacting protein 1-like to prevent metastasis of triple-negative breast cancer via sponging miR-300. Cancer Biomarkers, 39(2), 79-94.

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Colony Formation Assay Protocol

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For colony formation assays, approximately 600–800 cells per well were seeded on 6-well plates for separate interventions for 48 h. Then the medium was replaced every two days. After 10 days, cells were fixed in paraformaldehyde and stained with 0.2% crystalline violet (Beyotime, Shanghai, China).

Li J., Chen Q., Shi D, & Lian X. (2022). Combined time-restricted feeding and cisplatin enhance the anti-tumor effects in cisplatin-resistant and -sensitive lung cancer cells. Medical Oncology (Northwood, London, England), 40(1), 63.

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