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Colcemid

Manufactured by Cayman Chemical
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Colcemid is a lab reagent used for mitotic arrest. It functions by inhibiting microtubule polymerization, leading to the disruption of the mitotic spindle and subsequent arrest of cells in metaphase.

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Lab products found in correlation

Chloroquine diphosphate, by Merck Group (2 mentions) Mg132, by Selleck Chemicals (2 mentions) Axio imager a1 microscope, by Zeiss (1 mentions) Camostat mesylate, by Merck Group (1 mentions) Furin, by R&D Systems (1 mentions) Thermolysin, by Merck Group (1 mentions) Cytotox 96 non radioactive cytotoxicity colorimetric assay, by Promega (1 mentions) Aprotinin, by Cayman Chemical (1 mentions) Sb412515, by Cayman Chemical (1 mentions) Trypsin, by Merck Group (1 mentions) Nh4cl, by Merck Group (1 mentions) Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions) Superfrost plus slide, by Thermo Fisher Scientific (1 mentions) Sp5 inverted confocal microscope, by Leica (1 mentions) Cell recovery solution, by Corning (1 mentions) Glacial acetic acid, by Merck Group (1 mentions) Sodium citrate, by Merck Group (1 mentions) Ammonium chloride nh4cl, by Merck Group (1 mentions) Octadecyl rhodamine b chloride r18, by Thermo Fisher Scientific (1 mentions) Atto647n, by ATTO-TEC (1 mentions)

4 protocols using colcemid

1

Metaphase Chromosome Counting in F9 Cells

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Early- and late-passage F9 cells were cultured on 0.1% gelatin-coated coverslips and allowed to reach 80% confluency. Cells were treated with 0.2 μg/ml colcemid (Cayman Chemical) for 2 h at 37 °C, followed by trypsinization and suspension in pre-warmed 0.075 M KCl solution for 20 mins at 37 °C. Cells were fixed in acetic acid-methanol solution and then transferred onto pre-chilled glass slides. Chromosomes were stained with 4′,6-diamidino-2-phenylindole mounting media containing ProLong™ Gold Antifade Mountant and examined using an Axio Imager A1 microscope (Carl Zeiss). Thirty representative images were taken of each cell and chromosomes were counted manually for statistical analysis.
Gatie M.I, & Kelly G.M. (2018). Metabolic profile and differentiation potential of extraembryonic endoderm-like cells. Cell Death Discovery, 4, 42.
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2

Inhibitors of SARS-CoV-2 Cellular Entry

Cited 1 time
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Aprotinin (Cayman Chemical), camostat mesylate (Sigma), chloroquine diphosphate (Sigma), and NH4Cl (Sigma) stocks were dissolved in water. Bafilomycin A1 (BioViotica), colcemid (Cayman Chemical), concanamycin B (BioViotica), MG‐132 (Selleck Chemicals), SB412515 (Cayman Chemical), and DL‐threo‐1‐phenyl‐2‐palmitoylamino‐3‐morpholino‐1‐propanol (PPMP, Cayman Chemical) were all dissolved in DMSO. All drugs were assessed for cytotoxicity at the indicated concentrations using the CytoTox96 Non‐Radioactive Cytotoxicity colorimetric assay (Promega) according to the provider’s recommendations. Furin was purchased from R&D, and thermolysin and trypsin were purchased from Sigma. Plasmids encoding EGFP‐tagged Rab7a, Rab7a T22N, and Rab7a Q67L have been described elsewhere (Lozach et al, 2010 (link)).
Koch J., Uckeley Z.M., Doldan P., Stanifer M., Boulant S, & Lozach P. (2021). TMPRSS2 expression dictates the entry route used by SARS‐CoV‐2 to infect host cells. The EMBO Journal, 40(16), e107821.
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3

Metaphase Spread Preparation from Embryos

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Embryos were cultured in 0.1 µg ml−1 colcemid (Cayman chemical) for 12 h to arrest them in metaphase. Hypotonic solution: 1% sodium citrate (Sigma-Aldrich) and Carnoy’s fixative: 3:1 methanol: glacial acetic acid (Sigma-Aldrich). Eight- to 16-cell embryos were incubated for 10 min in pre-warm hypotonic solution and fixed for 30 min in Carnoy’s fixative. ICMs embedded in Matrigel were incubated in Cell Recovery Solution (Corning) for 20 min at 4 °C and then taken out of Matrigel using pipetting. They were incubated for 10 min in pre-warm hypotonic solution and fixed for 30 min in Carnoy’s fixative.
Embryos were mouth pipetted on clean SuperFrost Plus Slide (Thermo Fisher Scientific). Slides were air-dried and mounted with ProLong® Gold Antifade Mountant with DAPI (Life Technologies). Spreads were analysed using Leica SP5 inverted confocal microscope.
Singla S., Iwamoto-Stohl L.K., Zhu M, & Zernicka-Goetz M. (2020). Autophagy-mediated apoptosis eliminates aneuploid cells in a mouse model of chromosome mosaicism. Nature Communications, 11, 2958.
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4

Fluorescent Labeling of Endocytic Proteins

Cited 2 times
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The plasmids encoding Rab5a, Rab7a, and LAMP1 wt and mutant molecules tagged with EGFP have all been described previously [19 (link)]. Stock solutions of chloroquine diphosphate (Sigma) and ammonium chloride (NH4Cl, Sigma) were prepared in dH2O at concentrations of 19 mM and 1 M, respectively. Bafilomycin A1 (BioViotica), colcemid (Cayman Chemical), concanamycin B (BioViotica), MG-132 (Selleck Chemicals), nocodazole (Merck), and taxol (Merck) were all dissolved in 100% dimethyl sulfoxide (DMSO) to prepare stock solutions at 100 μM, 10 mM, 50 μM, 40 mM, 20 mM, and 10 mM, respectively. Stock solutions of all drugs were diluted in DMEM at the indicated doses (Figs 4 and 5), which are known not to cause cytotoxicity [23 (link), 34 (link)]. Both dH2O and DMSO were included as solvent controls. The hydroxysuccinimidyl (NHS) ester dyes AF488 (Thermo Fisher Scientific) and ATTO647N (Atto-Tec) were dissolved in DMSO (10 and 5 mg.mL-1, respectively) while octadecyl rhodamine B chloride (R18, Thermo Fisher Scientific) was dissolved in ethanol (10 mM).
Koch J., Xin Q., Obr M., Schäfer A., Rolfs N., Anagho H.A., Kudulyte A., Woltereck L., Kummer S., Campos J., Uckeley Z.M., Bell-Sakyi L., Kräusslich H.G., Schur F.K., Acuna C, & Lozach P.Y. (2023). The phenuivirus Toscana virus makes an atypical use of vacuolar acidity to enter host cells. PLOS Pathogens, 19(8), e1011562.
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