Karyomax colcemid solution in pbs

Manufactured by Thermo Fisher Scientific

KaryoMAX® Colcemid™ Solution in PBS is a laboratory reagent used for cell culture applications. It contains colcemid, a chemical compound that disrupts microtubule formation, leading to cell cycle arrest at metaphase. This solution is provided in a phosphate-buffered saline (PBS) matrix.

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Lab products found in correlation

Gurr s buffer, by Thermo Fisher Scientific (1 mentions) Giemsa solution, by Thermo Fisher Scientific (1 mentions) Calf serum, by Merck Group (1 mentions) Gentamicin, by Thermo Fisher Scientific (1 mentions) M5650, by Merck Group (1 mentions) Karyomax giemsa stain solution, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Amphotericin b, by Merck Group (1 mentions) Ultrakitt, by Avantor (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions)

3 protocols using karyomax colcemid solution in pbs

Chromosome Aberration Analysis of Hypoxic Cells

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Cells were synchronized into the G1 phase by mitotic shake off and 2 h of incubation. After ⁶⁴Cu-ATSM was added, cells were incubated for 3 h under hypoxic conditions, then 0.1 μg/ml Colcemid (KaryoMAX® Colcemid™ Solution in PBS, Gibco) was added and incubated for 12–16 h under normoxic conditions. Since we observed a wide variety in a degree of chromosome aberrations after ⁶⁴Cu-ATSM exposure, we assumed faster entry of less damaged cells and slower entry of heavily damaged cells to mitosis. This prolonged Colcemid treatment was aimed to collect a maximum population in metaphase cells. Harvested cells were treated with hypotonic solution (75 mM KCl) for 20 min at 37°C and fixed with methanol:acetic acid (3:1) solution three times before being dropped onto slides. Samples were stained with 5% (v/v) Giemsa solution in Gurr's buffer (Gibco). At least 50 metaphase cells were scored in at least two separate experiments. Chromosomal aberrations were quantified and classified as various chromosome and chromatid type aberrations.

McMillan D.D., Maeda J., Bell J.J., Genet M.D., Phoonswadi G., Mann K.A., Kraft S.L., Kitamura H., Fujimori A., Yoshii Y., Furukawa T., Fujibayashi Y, & Kato T.A. (2015). Validation of 64Cu-ATSM damaging DNA via high-LET Auger electron emission. Journal of Radiation Research, 56(5), 784-791.

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Preparation of Chromosome Spreads from Corn Snake Fibroblasts

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Chromosome spreads were prepared from corn snake fibroblasts established from an E30 embryo. Fibroblasts were grown at 30 °C in MEM (M5650; Sigma-Aldrich) supplemented with 2 mM L-glutamine (no. 25030024; Gibco), 10% calf serum (C8056; Sigma-Aldrich), 100 U/mL penicillin/streptomycin (no. 15140122; Gibco), 2.5 μg/mL amphotericin B (A2942; Sigma-Aldrich), and 50 μg/mL gentamicin (no. 15750037; Gibco). KaryoMAX Colcemid Solution in PBS (no. 15212012; Gibco) was used at 0.1 μg/mL to arrest cells in metaphase. Cells were collected after trypsin digestion and incubated for 10 min at 37 °C in a 0.075 M KCl solution. They were fixed in methanol:acetic acid (3:1), and the cell suspensions were dropped onto clean glass slides and air-dried. Chromosomes were stained with KaryoMAX Giemsa Stain Solution (no. 10092013; Gibco), rinsed with distilled water, air-dried, and mounted with Ultrakitt (no. 3921; JT Baker).

Ullate-Agote A., Burgelin I., Debry A., Langrez C., Montange F., Peraldi R., Daraspe J., Kaessmann H., Milinkovitch M.C, & Tzika A.C. (2020). Genome mapping of a LYST mutation in corn snakes indicates that vertebrate chromatophore vesicles are lysosome-related organelles. Proceedings of the National Academy of Sciences of the United States of America, 117(42), 26307-26317.

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Chromosome Preparation for BonFIRE Imaging

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HeLa cells were seeded in a 35-cm dish and labelled with EdU. Cells were incubated with 1 μg/mL colcemid (KaryoMAX^™ Colcemid^™ Solution in PBS, Gibco) for 4 hours. Cells were trypsinized into single cells with Trypsin-EDTA (0.25%, Gibco). To stop the digestion, 5 mL DMEM containing 10% FBS was added. The cell suspensions were centrifuged at 120 × g for 5 min and the supernatant was aspirated with about 200 μL left in the tube. Clumps were removed by gently tapping the bottom of the tube. 5 mL of ice-cold 0.56% KCl solution was added to the cell suspensions gently. After putting it at room temperature for 10 min, the cell suspensions were centrifuged at 120 × g for 5 min and the supernatant was aspirated with about 200 μl left in the tube. 5 ml of methanol: glacial acetic acid (3:1) fixative solution was added to the cell suspensions gently and slowly. The cell suspensions were centrifuged at 120 × g for 5 min and the cells were resuspended with PBS, after aspirating the supernatant. The cells were then clicked-labelled using the procedure described above. The clicked chromosomes were deposited onto a CaF₂ window and were dried for BonFIRE imaging.

McPhee F., Good A.C., Kuntz I.D, & Craik C.S. (1996). Engineering human immunodeficiency virus 1 protease heterodimers as macromolecular inhibitors of viral maturation.. Proceedings of the National Academy of Sciences of the United States of America, 93(21), 11477-11481.

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