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Aphidicolin

Most cited protocols related to «Aphidicolin»

Cited 53 times

HeLa, HCC1937, U2OS and U2OS-derived cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% foetal bovine serum (FBS) and standard antibiotics. Data for survival curves were generated by colony formation assays. In brief, U2OS cells were transfected with siRNA (see Supplementary Methods) and treated with DNA-damage-inducing drugs. After 1 h, the drug was removed and cells were left for 10-14 days at 37°C to allow colonies to form. Colonies were stained with 0.5% crystal violet/20% ethanol and counted. Where indicated, cells were pre-incubated with aphidicolin (10 μM) for 90 min, then treated with the specified drug and aphidicolin for 1 h. A U2OS-derived cell line stably expressing GFP-ATR was described previously7 (link). The siRNA-resistant silent wild-type GFP-CtIP construct was generated by sub-cloning the CtIP cDNA into the pEGFP-C1 expression plasmid (BD Biosciences Clontech) and changing three nucleotides in the CtIP-1 siRNA targeting region by using a QuikChange site-directed mutagenesis kit (Stratagene, Inc.) as previously described16 (link). The plasmid expressing the CtIP mutant lacking the C-terminus (1-789) was generated by changing residues 790 and 791 in the wild-type GFP-CtIP construct to two stop-codons. For generation of cell lines stably expressing siRNA-resistant GFP-tagged wild-type and mutant CtIP, U2OS cells were transfected with the appropriate constructs and, following antibiotic selection, resistant clones were tested for expression and nuclear localization of the transgene-product by immunofluorescence microscopy. To detect ssDNA by microscopy, cells were cultivated for 24 h in medium supplemented with 10 μM BrdU prior to camptothecin treatment and, after fixation, immunostained with an anti-BrdU antibody (see Methods) without any preceding DNA denaturation or nuclease treatment48 (link). Laser micro-irradiation was performed as described previously30 (link),31 (link). Recombinant FLAG-GST-CtIP-6H was isolated from baculovirus-infected Sf9 cells as described previously49 (link). Recombinant MRE11-RAD50 (MR) and MRE11-RAD50-NBS1 (MRN) complex were kind gifts from T. Paull.
Full Methods and any associated references are available in the online version of the paper at www.nature.com/nature.

Sartori A.A., Lukas C., Coates J., Mistrik M., Fu S., Bartek J., Baer R., Lukas J, & Jackson S.P. (2007). Human CtIP promotes DNA end resection. Nature, 450(7169), 509-514.

Publication 2007

Antibiotics Antibodies, Anti-Idiotypic Aphidicolin Baculoviridae Biological Assay Bromodeoxyuridine Camptothecin Cell Lines Cells Clone Cells Codon, Terminator DNA, Complementary DNA, Single-Stranded DNA Damage DNA Denaturation Eagle Ethanol Fetal Bovine Serum Gifts HeLa Cells Immunofluorescence Microscopy Microscopy Mutagenesis, Site-Directed Nucleotides Pharmaceutical Preparations Plasmids Rad50 protein, human Radiotherapy RBBP8 protein, human RNA, Small Interfering Sf9 Cells Transgenes Violet, Gentian

Astrocyte Survival Assay Protocol

Cited 14 times

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Publication 2011

Aphidicolin Astrocytes Cells Culture Media, Conditioned Endothelial Cells Growth Factor neuro-oncological ventral antigen 2, human Pericytes

Chromatid Break and SCE Analysis

Cited 12 times

For PCC analysis, cells were treated with 50 ng/ml calyculin A (Calbiochem) for 30 min before harvesting. For SCE analysis, cells were grown for 48 h in BrdU before irradiation. Colcemid (together with 1 mM caffeine to overcome the G2/M arrest) was added at 8 h until 12 h post-irradiation to collect cells in mitosis. Chromatid breaks (for PCC analysis) or SCEs were scored in at least 100 chromosome spreads from at least three independent experiments per data point. Staining was according to standard protocols. Aphidicolin (Sigma) was added at 1 μg/ml immediately before IR. ATM inhibitor (KU55933) and DNA-PK inhibitor (NU7026), a kind gift from Kudos Pharmaceuticals (Cambridge, UK), were added at 20 μM 60 min prior to IR. X-ray irradiation was performed at 90- or 120-kV γ-irradiation using a ¹³⁷Cs source. Dosimetry was performed with ion chambers and considered the increase in dose for cells grown on glass coverslips relative to plastic surfaces (Kegel et al, 2007 (link)).

Beucher A., Birraux J., Tchouandong L., Barton O., Shibata A., Conrad S., Goodarzi A.A., Krempler A., Jeggo P.A, & Löbrich M. (2009). ATM and Artemis promote homologous recombination of radiation-induced DNA double-strand breaks in G2. The EMBO Journal, 28(21), 3413-3427.

Publication 2009

Aphidicolin Bromodeoxyuridine Caffeine calyculin A Cells Chromatids Chromosomes Colcemide KU 55933 Mitosis NU 7026 Pharmaceutical Preparations Protein Kinase, DNA-Dependent Radiometry Radiotherapy X-Rays, Diagnostic

Targeted RNAi for Studying Nuclear Transport Factors

Cited 11 times

All RNA interference experiments were performed by expressing short hairpin RNA from either MLV vector pSIREN RetroQ (Clontech) (for Nup358, TRN-SR2 and Nup153) or pSUPER (Oligoengine) (for CypA) or if indicated from the HIV-1 vector pCSRQ, which was derived by subcloning the shRNA expression cassette from pSIREN RetroQ into pCSGW. The CypA shRNA target sequence has been described [58] (link). The Nup358 shRNA target sequence that was used throughout the study was 5-GCGAAGTGATGATATGTTT-3. Nup153 shRNA target sequence was 5-CAATTCGTCTCAAGCATTA-3. Both sequences were selected as 1 of the 4 target sequences from the Dharmacon siRNA smartpool for Nup358 or Nup153, respectively. Both shRNAs had only minor toxic effects on the cells, unlike shRNAs derived from the other three target sequences of each smart pool (Figure S1A, and data not shown). Additional Nup358 shRNA target sequences used in the experiment shown in Figure S1A were shRNA2 5-CAAACCACGTTATTACTAA-3, shRNA3 5-CAGAACAACTTGCTATTAG-3 and shRNA4 5-GAAGGAATGTTCATCAGGA-3. Specificity for each target sequence was confirmed by BLAT (UCSC genome browser). For Nup358, we confirmed effective targeting by co-transfecting the shRNA expression vector with a plasmid encoding GFP-tagged Nup358 (Figure S2), as well as by western blotting using a Nup358 specific antibody (Figure 1B). TRN-SR2 target sequence and control have been described and were also validated by co-transfecting a plasmid encoding for TRN-SR2-IRES-eGFP with the expression vectors encoding shRNA or control (SC) (Figure S2) [22] (link). The observations made for shRNA expressing HeLa cells were similar between populations of puromycin selected cells and clonal cells but the phenotype of cell clones was more stable, thus we used single cell clones for all experiments (Figure S1A, B and data not shown). Nup358, TRN-SR2, Nup153, CypA and beta-Actin were detected by western blot using a Nup358 antibody kindly given by Frauke Melchior, mouse TRN-SR2 antibody ab54353 (Abcam), mouse Nup153 antibody ab24700 (Abcam), rabbit CypA antibody SA296 (Biomol) and mouse beta-Actin antibody ab6276 (Abcam) and appropriate horseradish peroxidase linked secondary antibodies. TRIMCypA and TRIMNup358 were detected using anti-HA antibody 3F10 (Roche). Cyclosporine (Sandoz) and aphidicolin (Sigma) were diluted in DMSO and used at 5–8 µM and 2 µg/ml, respectively.

Schaller T., Ocwieja K.E., Rasaiyaah J., Price A.J., Brady T.L., Roth S.L., Hué S., Fletcher A.J., Lee K., KewalRamani V.N., Noursadeghi M., Jenner R.G., James L.C., Bushman F.D, & Towers G.J. (2011). HIV-1 Capsid-Cyclophilin Interactions Determine Nuclear Import Pathway, Integration Targeting and Replication Efficiency. PLoS Pathogens, 7(12), e1002439.

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Publication 2011

Antibodies Antibodies, Anti-Idiotypic Aphidicolin beta-Actin Cells Clone Cells Cloning Vectors Cyclosporine Genome HeLa Cells HIV-1 Horseradish Peroxidase Immunoglobulins Internal Ribosome Entry Sites Mus Phenotype Plasmids Population Group Puromycin Rabbits RNA, Small Interfering RNA Interference Short Hairpin RNA Sulfoxide, Dimethyl Western Blotting

Immunofluorescence Detection of G-Quadruplexes

Cited 9 times

U2OS (osteosarcoma), HeLa (cervical carcinoma), HT1080 (fibrosarcoma), MCF-7 (mammary adenocarcinoma) and MDA-MB-231 (breast carcinoma) cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Gibco), 1% L-glutamine, 10% FBS (fetal bovine serum); MRC-5 (fetal lung fibroblasts) cells in Minimum Essential Medium Eagle (Sigma), 10% FBS and MCF-10A (mammary epithelial) cells in Mammary Epithelial cell Growth Medium (Lonza) with cholera toxin (at 37 °C with 5% CO₂. Cells grown on glass coverslips were fixed in 2% paraformaldehyde/PBS or in methanol: acetic acid (3:1) (Supplementary Fig. S3) and permeabilized with 0.1% triton-X100/PBS. After blocking in 2% Marvel™/PBS, immunofluorescence was performed using standard methods with BG4, anti-FLAG (#2368, Cell Signaling Technology) and anti-rabbit Alexa 594-conjugated (A11037, Invitrogen) antibodies. Coverslips were mounted with Prolong Gold/DAPI (4′,6-diamidino-2-phenylindole) (Invitrogen). For TRF2 detection, anti-TRF2 (ab13579, Abcam) and anti-mouse Alexa 488-conjugated (A11029, Invitrogen) antibodies were used. For enzyme treatments, coverslips were incubated after permeabilization with 120 U of Turbo DNase (0.12 U/μl) or 50 μg/ml of RNase A (Ambion) for 1 h at 37 °C. For pre-incubation experiments, BG4 was incubated with 20-fold excess of pre-folded DNA G-quadruplexes or single-stranded DNA oligonucleotides. Oligonucleotide transfections were performed using 200 nM pre-annealed DNA G-quadruplexes or single-stranded DNA oligonucleotides and TransIT™ Oligo Transfection Reagent (Mirus). For pyridostatin treatment, cells were incubated with 10 μM compound for 24 h. For metaphase preparations, cells were treated with colcemid (50 ng/ml) for 2 h before resuspension in 0.075 mM KCl for 30 min at 37 °C. Cells were then fixed in methanol: acetic acid (3:1) before spreading on slides. 100 well-spread metaphase chromosomes were analyzed for the number and localization of BG4 foci. For cell synchronization, MCF-7 cells were incubated for 24 h in serum-free DMEM (G0/G1), grown for 16 h in DMEM, 20% FBS, 200 μM mimosine (G1/S), and for 3 h in DMEM, 10% FBS (S phase). Cell cycle stages were confirmed using a fluorescent activated cell sorter (FACSCalibur, BD Biosciences). To inhibit DNA replication, cells were incubated with 5 μM aphidicolin for 2 h. Digital images were recorded using a DP70 camera (Olympus) on Axioskop 2 plus microscope (Zeiss), and analyzed with Volocity software (Perkin Elmer). 100-200 nuclei were counted per condition and the standard error mean calculated from 3 replicates. Frequency distribution graphs were plotted using GraphPad Prism (GraphPad Software, Inc.).

Biffi G., Tannahill D., McCafferty J, & Balasubramanian S. (2013). Quantitative Visualization of DNA G-quadruplex Structures in Human Cells. Nature chemistry, 5(3), 182-186.

Publication 2013

Acetic Acid Adenocarcinoma Alexa594 Antibodies Aphidicolin Breast Breast Carcinoma Cardiac Arrest Cell Cycle Cell Nucleus Cells Cervical Cancer Cholera Toxin Chromosomes Colcemide DAPI Deoxyribonucleases DNA, Single-Stranded DNA Replication Eagle Endoribonucleases Enzymes Epithelial Cells Fetus Fibroblasts Fibrosarcoma G-Quadruplexes, DNA Glutamine Gold HeLa Cells Immunofluorescence Lung MCF-7 Cells Metaphase Methanol Microscopy Mimosine Mus Oligonucleotides Osteosarcoma paraform prisma pyridostatin Rabbits Serum Transfection Triton X-100

Most recents protocols related to «Aphidicolin»

Isolation and culture of various cell types

MEFs were maintained as previously published (Hall et al., 2013 (link)). SNAP labeling was performed as previously described (Quidwai et al., 2021 (link)). Ependymal cells were isolated and cultured as published in Delgehyr et al., 2015 (link). mTECs were isolated and cultured as described in Eenjes et al., 2018 (link); You et al., 2002 (link). RPE1-hTERT (female, human epithelial cells immortalized with hTERT, Cat. No. CRL-4000) from ATCC were grown in Dulbecco's Modified Eagle Medium (DMEM, Life Technologies) or DMEM/F12 (Thermo Fisher Scientific, 10565042) supplemented with 10% fetal bovine serum at 37°C with 5% CO₂. For live imaging, the membrane was cut out and placed cilia down on a glass dish (Nest, 801002) in a drop of media. PCM1^−/− RPE1 cells were generated as described previously (Kumar et al., 2021 (link)) (all figures except for Figure 8—figure supplement 1, in which case they were generated as in Gheiratmand et al., 2019 (link)). hTERT-RPE1: Source ATCC, confirmed mycoplasma negative and verified by STR profiling. Two PCM1^−/− RPE1 cell lines were generated using single guide RNAs (Supplementary file 1). Loss of PCM1 was confirmed by genotyping, immunoblotting, and immunofluorescence. Monoclonal PCM1^−/− RPE1 cell lines stably expressing eGFP or eYFP-PCM1 (plasmid a gift from Bryan Dynlacht; Wang et al., 2016 (link)) were generated using lentiviruses and manually selected based on fluorescence. To synchronize cells in G1/S aphidicolin (Sigma) was added to the culture medium at 2 μg/ml for 16 hr. To arrest cells in mitosis, taxol (paclitaxel; Millipore-Sigma) was added to the culture medium at 5 μM for 16 hr prior to rounded up cells being collected by mitotic shake-off. For arrest in G0, cells were washed 2× with phosphate-buffered saline (PBS; Gibco) and 1× with DMEM (without serum) before being cultured in serum-free DMEM for 16 hr. To disrupt cytoplasmic microtubules, cells were treated with 20 μM nocodozole (Sigma, SML1665) for 1–2 hr prior to fixation.

Hall E.A., Kumar D., Prosser S.L., Yeyati P.L., Herranz-Pérez V., García-Verdugo J.M., Rose L., McKie L., Dodd D.O., Tennant P.A., Megaw R., Murphy L.C., Ferreira M.F., Grimes G., Williams L., Quidwai T., Pelletier L., Reiter J.F, & Mill P. (2023). Centriolar satellites expedite mother centriole remodeling to promote ciliogenesis. eLife, 12, e79299.

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Publication 2023

Aphidicolin Cardiac Arrest Cell Lines Cells Cilia Culture Media Cytoplasm Dietary Supplements Eagle Ependyma Epithelial Cells Females Fetal Bovine Serum Fluorescence Homo sapiens Hyperostosis, Diffuse Idiopathic Skeletal Immunofluorescence Lentivirus Microtubules Mitosis Mycoplasma Paclitaxel Phosphates Plasmids RNA, Single Guide Saline Solution Serum Taxol Tissue, Membrane Tremor

Probing ORC1 Protein Regulation

To study the regulation of the ORC1 proteins, GFP-tagged seedlings were incubated in liquid MS media containing different inhibitors. Plants (4 days post sowing; dps) were acclimated in the liquid MS media during ~12 h to recover from the temporary hypoxia. To inhibit the proteasome pathway plants were incubated with 50 µM bortezomib (Selleckchem) for 4 h or with 250 µM MLN4924 (APExBIO) for 6 h. Control plants were incubated in MS containing DMSO used as the drugs solvent. Incubations were carried out in the plant culture chamber. To assess root growth under DNA replication stress conditions, 3 dps orc1 mutants, orc1b-2,ORC1b-GFP and Col-0 seeds were grown in 1% agar MSS plates and transferred to 1% agar MS plates containing 0.5% sucrose supplemented with 1 mM hydroxyurea (Sigma), 12 µg/ml aphidicolin (Sigma) or no drug as controls. Seeds that did not germinate were removed from the analysis. Root growth was measured every 24 h for a total of 10 days.

Vergara Z., Gomez M.S., Desvoyes B., Sequeira-Mendes J., Masoud K., Costas C., Noir S., Caro E., Mora-Gil V., Genschik P, & Gutierrez C. (2023). Distinct roles of Arabidopsis ORC1 proteins in DNA replication and heterochromatic H3K27me1 deposition. Nature Communications, 14, 1270.

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Publication 2023

Agar Aphidicolin Bortezomib Cardiac Arrest DNA Replication Hydroxyurea Hypoxia inhibitors MLN4924 Multicatalytic Endopeptidase Complex Origin Recognition Complex, Subunit 1 Pharmaceutical Preparations Plant Embryos Plant Roots Plants Seedlings Solvents Stress Disorders, Traumatic Sucrose Sulfoxide, Dimethyl

DNA Damage Inducers and Replication Inhibitors

(i) DNA damage inducer and PI3KK inhibitors. Hydroxyurea (HU; #127-07-1; MilliporeSigma) was prepared as a stock solution at 200 mM, according to manufacturer’s instructions. PI3KK inhibitors, KU60019 (#4176; Tocris Bioscience), AZ20 (#S7050; Selleckchem), and NU7441 (#3712; Tocris Bioscience), were dissolved in DMSO at 10 mM. They were applied to cell cultures at 2 days prior to infection at the final concentrations, as we previously reported: HU at 5 mM, KU60019 at 5 μM, AZ20 at 3 μM, and NU7441 at 1 μM (31 (link), 32 (link), 80 (link)).
(ii) In vitro DNA replication inhibitors. We used HAMNO (#S0148, Selleckchem), T2AA (#21921, Cayman Chemical), aphidicolin (#14007, Cayman Chemical), MK886 (#1311, Tocris Bioscience), and PNR7-02 (#2965, Axon Medchem). Stock solutions were prepared in DMSO at 10 mM. The inhibitors were directly added into the reaction of in vitro replication assays at a final concentration of 100 μM for HAMNO, 1 μM for T2AA, 10 μg/μL for aphidicolin, 200 μM for MK886, and 20 μM for PNR7-02 as previously recommended (44 (link)– (link)48 (link)).

Ning K., Kuz C.A., Cheng F., Feng Z., Yan Z, & Qiu J. (2023). Adeno-Associated Virus Monoinfection Induces a DNA Damage Response and DNA Repair That Contributes to Viral DNA Replication. mBio, 14(1), e03528-22.

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Publication 2023

Aphidicolin Axon Biological Assay Caimans Cell Culture Techniques DNA Damage DNA Replication DNA Synthesis Inhibitors Hydroxyurea Infection inhibitors KU 60019 NU 7441 Sulfoxide, Dimethyl T2AA

MS-BAND Protocol for DNA Replication Dynamics

Fig. S1 shows material related to Fig. 1 (LC-MS/MS chromatograms of nucleoside standards, LC and MS characteristics of nucleosides, quantitative standard curves, and detection of nascent DNA using MS-BAND). Fig. S2 shows material related to Fig. 2 (MS-BAND based quantitation of thymidine analogue incorporation after DNA polymerase inhibition with aphidicolin). Fig. S3 shows material related to Fig. 4 (mtDNA purification, IdU and CldU incorporation into mtDNA as determined by MS-BAND, validation of PolG phenotype using slot blotting, and mito-MS-BAND phenotype of cells treated with EtBr). Fig. S4 is related to Fig. 5 (IdU and CldU incorporation into bacterial genomic DNA as determined by MS-BAND, and optical densities of control bacteria and temperature-sensitive mutant strains). Fig. S5 is related to Fig. 6 (validation of MS-BAND results in bacterial strains using a shorter thymidine analog incubation time and lack of correlation between MS-BAND and specific markers of DNA damage responses in bacteria). Table S1 is related to Fig. 6 (Z-scores of bacterial MS-BAND screen).

Ashour M.E., Byrum A.K., Meroni A., Xia J., Singh S., Galletto R., Rosenberg S.M., Vindigni A, & Mosammaparast N. (2023). Rapid profiling of DNA replication dynamics using mass spectrometry–based analysis of nascent DNA. The Journal of Cell Biology, 222(4), e202207121.

Publication 2023

Aphidicolin Bacteria DNA, Bacterial DNA, Mitochondrial DNA-Directed DNA Polymerase EDNRB protein, human Genome Markers, DNA Mitomycin Neutrophil Band Cells Nucleosides Phenotype POLG protein, human Psychological Inhibition Strains Tandem Mass Spectrometry Thymidine Vision

Studying DNA Replication Stress in Mitosis

To introduce replication stress, cells were treated with 0.4 μM aphidicolin for the period indicated in the figures. To induce POLD1 degradation, a modified HCT116 cell line that has endogenous POLD1 targeted by CRISPR-Cas9 and exogenous stable POLD1-OsTIR1(F74G) expression (the HCT116-POLD1-AID2 cell line) was treated with 1 μM 5-Ph-IAA (BioAcademia) for the periods indicated in the Fig. 1a–e.
To enrich mitotic cells, asynchronous cells were treated with CDK1 inhibitor RO-3306 (7 μM; APExBIO) for 6 h, or in the last 6 h of APH treatment where necessary. Cells were then rinsed with pre-warmed medium (37 °C) for three times (within 5 min) before being released into fresh cell culture (with EdU, or other relevant treatment) to allow them to progress into mitosis for the follow-up analyses.
For MiDAS analysis in prophase/prometaphase, EdU was added in the pre-warmed medium (37 °C) and incubated with cells for 30 min. Prometaphase cells were then collected by mitotic-shake off, re-seeded onto Poly-L-Lysine-coated slides (Sigma Aldrich) for IF analysis, or collected by centrifugation for Western blotting (WB) analysis. To obtain anaphase cells, prometaphase cells collected from mitotic-shake off were re-seeded onto pre-warmed, Poly-L-Lysine-coated slides (Sigma Aldrich), and were incubated for additional 20 min to allow them to progress into anaphase for anaphase bridges analysis. To obtain G1 phase cells for micronuclei and 53BP1 foci analysis, prometaphase cells collected from mitotic-shake off were re-seeded onto pre-warmed, Poly-L-Lysine-coated slides and were then incubated for an additional 2.5 h to allow them to progress into the next G1 phase.

Wu W., Barwacz S.A., Bhowmick R., Lundgaard K., Gonçalves Dinis M.M., Clausen M., Kanemaki M.T, & Liu Y. (2023). Mitotic DNA synthesis in response to replication stress requires the sequential action of DNA polymerases zeta and delta in human cells. Nature Communications, 14, 706.

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Publication 2023

Anaphase Aphidicolin CDK1 protein, human Cell Culture Techniques Cell Lines Cells Centrifugation Clustered Regularly Interspaced Short Palindromic Repeats DNA Replication G1 Phase GOLPH3 protein, human HCT116 Cells Lysine Mitosis POLD1 protein, human Poly A Prometaphase RO 3306 TP53BP1 protein, human Tremor Western Blot

Top products related to «Aphidicolin»

Aphidicolin by Merck Group

Sourced in United States, Germany, United Kingdom, Denmark

Aphidicolin is a laboratory reagent that functions as a DNA polymerase inhibitor. It is commonly used in cell biology and molecular biology research applications.

Hydroxyurea by Merck Group

Sourced in United States, Germany, France, Italy, United Kingdom

Hydroxyurea is a chemical compound used in various laboratory applications. It functions as an inhibitor of ribonucleotide reductase, an enzyme involved in the synthesis of DNA.

Nocodazole by Merck Group

Sourced in United States, Germany, United Kingdom, Japan, Sao Tome and Principe, Canada, China, Switzerland, France, Poland, Macao, Australia

Nocodazole is a synthetic compound that acts as a microtubule-destabilizing agent. It functions by binding to and disrupting the polymerization of microtubules, which are essential components of the cytoskeleton in eukaryotic cells. This property makes Nocodazole a valuable tool in cell biology research for studying cell division, cell motility, and other cellular processes that rely on the dynamics of the microtubule network.

Thymidine by Merck Group

Sourced in United States, Germany, United Kingdom, Sao Tome and Principe, France, Italy, Japan, China

Thymidine is a nucleoside that is a component of DNA. It serves as a building block for DNA synthesis and is essential for cellular division and growth.

Aphidicolin by Cayman Chemical

Sourced in United States

Aphidicolin is a chemical compound used as a laboratory reagent. It functions as a reversible inhibitor of DNA polymerase α, which is responsible for the initiation of DNA replication. Aphidicolin is commonly used in cell biology research to synchronize cell populations in the G1/S phase of the cell cycle.

Dimethyl sulfoxide (dmso) by Merck Group

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DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.

Dulbecco modified eagle medium (dmem) by Thermo Fisher Scientific

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DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture medium formulated to support the growth and maintenance of a variety of cell types, including mammalian cells. It provides essential nutrients, amino acids, vitamins, and other components necessary for cell proliferation and survival in an in vitro environment.

Aphidicolin by Fujifilm

Sourced in Japan

Aphidicolin is a DNA polymerase inhibitor used in laboratory research. It selectively blocks the activity of DNA polymerase α, leading to the inhibition of DNA replication. Aphidicolin is commonly used as a tool to study cell cycle regulation and synchronize cells in the G1/S phase transition.

Camptothecin by Merck Group

Sourced in United States, Germany, United Kingdom, Belgium, Sao Tome and Principe, France, Macao, Poland

Camptothecin is a naturally occurring alkaloid compound isolated from the Camptotheca acuminata tree. It is a biologically active compound with potential applications in research and development. Camptothecin exhibits inhibitory effects on the enzyme topoisomerase I, which is involved in DNA replication and transcription processes. This property makes Camptothecin a valuable tool for studying cellular mechanisms and biological processes.

Lipofectamine 2000 by Thermo Fisher Scientific

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Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.

What is Aphidicolin and how is it commonly used in cell biology research?

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What are some common challenges or variations when working with Aphidicolin?

One challenge when working with Aphidicolin is ensuring the optimal dosage and timing to achieve the desired cell cycle synchronization without causing unintended effects. Additionally, there are different formulations and derivatives of Aphidicolin that may have slightly varied properties and applications, requiring researchers to carefully evaluate which version is best suited for their experiments.

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More about "Aphidicolin"

Aphidicolin is a potent DNA polymerase inhibitor derived from the fungus Cephalosporium aphidicola.
It is commonly used in cell biology research to synchronize cells in the G1/S phase transition, allowing the study of DNA replication and cell cycle regulation.
Aphidicolin is a valuable tool for researchers investigating the cell cycle, DNA synthesis, and related processes.
Hydroxyurea is another commonly used cell cycle synchronization agent that works by inhibiting ribonucleotide reductase, an enzyme essential for DNA synthesis.
Nocodazole, on the other hand, is a microtubule-disrupting agent that blocks cells in the M phase of the cell cycle.
Thymidine is a nucleoside that can also be used to synchronize cells, often in combination with Aphidicolin or other agents.
DMSO (Dimethyl sulfoxide) is a widely used solvent for dissolving and delivering various compounds, including cell cycle inhibitors, to cells.
DMEM (Dulbecco's Modified Eagle Medium) is a commonly used cell culture medium that provides the necessary nutrients and growth factors for cell proliferation and maintenance.
Camptothecin is a topoisomerase I inhibitor that can induce DNA damage and cell cycle arrest, while Lipofectamine 2000 is a transfection reagent often used to introduce genetic material into cells.
By utilizing the insights gained from these related terms and techniques, researchers can optimize their Aphidicolin-based experiments, enhancing the reproducibility and accuracy of their cell biology research.
PubCompare.ai can help streamline this process by providing easy access to the best experimental protocols and products from literature, pre-prints, and patents, taking the guesswork out of your Aphidicolin research.
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