Lt07 418

Manufactured by Lonza

Sourced in United States

The LT07-418 is a laboratory equipment product manufactured by Lonza. It is designed to perform a core function, but a detailed description while maintaining an unbiased and factual approach is not available.

Automatically generated - may contain errors

Lab products found in correlation

Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (5 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions) Cycloheximide (chx), by Merck Group (2 mentions) Phorbol myristate acetate (pma), by Merck Group (2 mentions) Mg132, by Merck Group (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Harvard Bioscience (2 mentions) Penicillin, by Lonza (2 mentions) Streptomycin, by Lonza (2 mentions) L glutamine, by Lonza (2 mentions) Genome wide human snp array 6, by Thermo Fisher Scientific (1 mentions) Ham s f12 media, by Thermo Fisher Scientific (1 mentions) Apo transferrin, by Merck Group (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Insulin, by Merck Group (1 mentions) Hepes, by Merck Group (1 mentions) Hydrocortisone, by Merck Group (1 mentions) Ethanolamine, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions)

14 protocols using lt07 418

Derivation and Authentication of UCLA hESC Lines

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The UCLA2 and UCLA1 hESC lines were derived at UCLA by the UCLA Pluripotent Stem Cell Core Facility following Institutional Review Board (IRB) and UCLA Embryonic Stem Cell Research Oversight (ESCRO) Committee Approvals. Informed consent was obtained after the embryo donors contacted the UCLA Broad Stem Cell Research Center to inquire about donating surplus embryos following in vitro fertilization. Embryo donors were not paid and were able to freely withdraw consent to use the embryos for stem cell research up to the point of hESC derivation when the embryo is destroyed. Informed consent was obtained from all embryo donors prior to sending frozen donated embryos to UCLA. Once derived, the hESC lines were authenticated using Affymetrix Genome-wide Human SNP Array 6.0 to detect Single Nucleotide Polymorphisms and Copy Number Variant (SNP/CNV) prior to distribution. The UCLA1 and UCLA2 hESC lines are provided to researchers de-identified, with all links and identifiers broken prior to distribution. All de-identified hESC lines used in this study are registered with the National Institute of Health Human Embryonic Stem Cell Registry and are available for research use with NIH funds. Mycoplasma test (Lonza, LT07-418) was performed every month. All experiments using the de-identified hESC lines were approved by the UCLA Embryonic Stem Cell Research Oversight Committee.

Xiang X., Tao Y., DiRusso J., Hsu F.M., Zhang J., Xue Z., Pontis J., Trono D., Liu W, & Clark A.T. (2022). Human reproduction is regulated by retrotransposons derived from ancient Hominidae-specific viral infections. Nature Communications, 13, 463.

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CRISPR-Mediated Depletion of RepID in K562 Cells

Cited 2 times

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Human K562 cells were cultured in Dulbecco’s modified Eagle’s medium (Invitrogen, 10569–010) supplemented with 10% heat-inactivated fetal bovine serum in a 37 °C/5% CO2 humidified incubator [19 (link), 21 (link)]. Original cell lines were obtained from American Type Culture Collection (ATCC) (www.atcc.org) and tested for mycoplasma (Lonza, LT07-418). Cycloheximide, MG132, and PMA were purchased from Sigma (Cat. C4859), Millipore (Cat. 11134515001), and Santa Cruz (Cat. Sc-3576), respectively. Each compound was added to the medium at the indicated concentrations.
RepID was depleted in K562 cells using CRISPR-CAS9 via 20-base pair guide sequence targeting the fifth exon of RepID (5’-CTGCAAATATGTCATCGACTAGG-3’) and the eighth exon of RepID (5’-GTGATAAAATGATCCGAGTCTGG-3’) from the published database of predicted high specificity protospacer-PAM target sites in the human exome. Cloning, selection and verification using PCR were performed as described previously [19 (link), 21 (link)].

Jo J.H., Park J.U., Kim Y.M., Ok S.M., Kim D.K., Jung D.H., Kim H.J., Seong H.A., Cho H.J., Nah J., Kim S., Fu H., Redon C.E., Aladjem M.I, & Jang S.M. (2023). RepID represses megakaryocytic differentiation by recruiting CRL4A-JARID1A at DAB2 promoter. Cell Communication and Signaling : CCS, 21, 219.

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Culturing IBC Cell Lines with Supplements

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All IBC cell lines were grown in HAMS F12 media (Gibco 11765–054) in a 5% CO₂ incubator. Media for SUM-149 cells was supplemented with 5% FBS (Atlanta Biologicals), 10mM HEPES (Thermo Fisher 15630080), 1x antibiotic-antimycotic (anti-anti, Thermo Fisher 15240062), 1μg/mL hydrocortisone (Sigma H4001), and 5μg/mL insulin (Sigma I9278). SUM-190 media was supplemented with 1% FBS, 1μg/mL hydrocortisone, 5μg/mL insulin (Sigma I0516), 50nM sodium selenite (Sigma S9133), 5μg/mL apo-Transferrin (Sigma T-8158), 10nM triiodo thyronine (T3, Sigma T5516), 10mM HEPES, and 0.03% ethanolamine (Sigma 411000). MDA-IBC-3 cells were grown with 10% FBS, 1μg/mL hydrocortisone, 1x anti-anti, and 5μg/mL insulin (Sigma I0516). SUM cell lines were obtained from Stephen Ethier at the Medical University of South Carolina, and MDA-IBC-3 cells were obtained directly from Wendy Woodward at the University of Texas MD Anderson Cancer Center. All cell lines were routinely tested for mycoplasma contamination (Lonza LT07–418) and were authenticated using fragment analysis at the University of Michigan DNA sequencing core. Olaparib (MedChem Express HY-10162) and veliparib (MedChem Express HY-10129) were reconstituted in 100% DMSO for cellular assays.

Michmerhuizen A.R., Pesch A.M., Moubadder L., Chandler B.C., Wilder-Romans K., Cameron M., Olsen E., Thomas D.G., Zhang A., Hirsh N., Ritter C.L., Liu M., Nyati S., Pierce L.J., Jagsi R, & Speers C. (2019). PARP1 Inhibition Radiosensitizes Models of Inflammatory Breast Cancer to Ionizing Radiation. Molecular cancer therapeutics, 18(11), 2063-2073.

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Cell Culture Conditions for Cancer Models

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MEFs and mouse breast tumor lines: DMEM (Corning mt10013cv) supplemented with 10% FBS (Atlanta Biologicals), 1% pen/strep (Fisher 30002ci) and 2mM L-glutamine (total 6mM) (Fisher MT25005CI). MDA-MB468, MDA-MB 231, Myc-CaP (2015), and U87: DMEM supplemented with 10% FBS and 1% pen/strep. HCC1419, HCC1187, HCC 1937, HCC 1806, BT549, ZR75-1, PC3, LNCAP, DBTRG: RPMI (Fisher 10040cv) supplemented with 10% FBS and 1% pen/strep. CaP8 cells (2015): DMEM with 10% FBS, 1% pen/strep, and 5ug/mL insulin (Sigma I9278). Neurospheres: stem cell media with 10ug/mL FGF (R&D Systems 233-FB-025), 20ug/mL EGF (Peprotech AF-100-15) and heparin (from Dr. Raymund Yong, May 2015). All cells were cultured in a 37°C incubator with humidity and 5% CO₂. Cell lines were obtained from ATCC (which authenticate cell lines using several methods including DNA fingerprinting) in 2006, with the exception of MEFs, MCCL-278, and MCCL-357 which were produced in our lab from mice (2012-2016). Cell lines were clear of mycoplasma as determined by the Lonza kit (LT07-418) within 6 months of their use. Cell lines were further authenticated in 2015 by LabCorp using a short tandem repeat method.

Mathur D., Stratikopoulos E., Ozturk S., Steinbach N., Pegno S., Schoenfeld S., Yong R., Murty V.V., Asara J.M., Cantley L, & Parsons R. (2017). PTEN regulates glutamine flux to pyrimidine synthesis and sensitivity to dihydroorotate dehydrogenase inhibition. Cancer discovery, 7(4), 380-390.

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Culturing Human Cell Lines with Pharmacological Treatments

Cited 2 times

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Human K562 and DMS114 cells, parental and RepID knockout, were cultured in Dulbecco’s modified Eagle’s medium (Invitrogen, 10569–010) or RPMI-1640 medium (Invitrogen, 11875–093) supplemented with 10% heat-inactivated fetal bovine serum in a 37°C/5% CO2 humidified incubator (16 (link), 18 (link)). All original cell lines were obtained from American Type Culture Collection (ATCC) (www.atcc.org) and tested for mycoplasma (Lonza, LT07–418). Cycloheximide, MG132, and PMA were purchased from Sigma (Cat. C4859), Millipore (Cat. 11134515001), and Santa Cruz (Cat. Sc-3576), respectively. Each compound was added to the medium at the indicated concentrations.

Jo J.H., Ok S.M., Kim D.K., Kim Y.M., Park J.U., Jung D.H., Kim H.J., Seong H.A., Cho H.J., Nah J., Kim S., Fu H., Redon C.E., Aladjem M.I, & Jang S.M. (2023). RepID represses megakaryocytic differentiation by recruiting CRL4A-JARID1A at DAB2 promoter. Research Square.

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Characterization of hESC Lines for Research

Cited 1 time

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The hESC lines in this study are as follows: UCLA1 (46, XX), UCLA2 (46, XY)(Diaz Perez et al., 2012 (link)), UCLA8 (46, XX)(Chen et al., 2017 (link)), and H1 OCT4-GFP (H1) (46, XY)(Gkountela et al., 2015 (link)). All hESCs were cultured on mitomycin C-inactivated mouse embryonic fibroblasts (MEFs) and split every 7 days using Collagenase type IV (GIBCO, 17104–019). hESC media was comprised of 20% knockout serum replacement (KSR) (GIBCO, 10828–028), 100mM L-Glutamine (GIBCO,25030–081), 1x MEM Non-Essential Amino Acids (NEAA) (GIBCO, 11140–050), 55mM 2-Mercaptoethanol (GIBCO, 21985–023), 10ng/mL recombinant human FGF basic (Proteintech HZ1285), 1x Penicillin-Streptomycin (GIBCO, 15140–122), and 50ng/mL primocin (InvivoGen, ant-pm-2) in DMEM/F12 media (GIBCO, 11330–032). All hESC lines used in this study are registered with the National Institute of Health Human Embryonic Stem Cell Registry and are available for research use with NIH funds. hESCs used in this study were routinely tested for mycoplasma (Lonza, LT07–418). All experiments were approved by the UCLA Embryonic Stem Cell Research Oversight Committee.

Hancock G., Liu W., Peretz L., Chen D., Gell J., Collier A., Zamudio J., Plath K, & Clark A. (2021). Divergent roles for KLF4 and TFCP2L1 in Naive Ground State Pluripotency and Human Primordial Germ Cell Development. Stem cell research, 55, 102493.

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Comprehensive Cell Line Culture Protocols

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Cell lines were obtained from lab stocks and identities were confirmed via short tandem repeat testing. Cells tested Mycoplasma negative by fluorescence and/or ATP assay (Lonza LT07-418). Cal51 (RRID CVCL_1110) and MDA-MB-231 (RRID CVCL_0062) cells were cultured in DMEM + 10% FBS, 2 mmol/L l-glutamine, and 50 μg/mL pen/strep. MCF7 (RRID CVCL_0031) cells were cultured in the same media + 10 μg/mL insulin. MCF10A cells (RRID CVCL_0598) were cultured in DMEM:F12 with 1% horse serum, 20 ng/mL EGF, 500 μg/mL hydrocortisone, 100 ng/mL cholera toxin, 10 μg/mL insulin, and 50 μg/mL pen/strep. T47D cells (RRID CVCL_YX85) were cultured in RPMI1640 medium with 10% FBS, 2 mmol/L l-glutamine, and 50 μg/mL pen/strep. Each line was maintained at 37°C and 5% CO₂.

Tucker J.B., Bonema S.C., García-Varela R., Denu R.A., Hu Y., McGregor S.M., Burkard M.E, & Weaver B.A. (2023). Misaligned Chromosomes are a Major Source of Chromosomal Instability in Breast Cancer. Cancer Research Communications, 3(1), 54-65.

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U2OS Cell Culture and Treatment

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Human U2OS cells with or without RepID/RepID mutants were incubated in Dulbecco’s modified Eagle’s medium (Invitrogen, 10569-010) supplemented with 10% heat-inactivated fetal bovine serum in a 37 °C/5% CO₂ humidified incubator. Original U2OS osteosarcoma cell lines were obtained from the American Type Culture Collection (ATCC; www.atcc.org). All cell lines tested negative for mycoplasmas (Lonza, LT07-418). MLN4924 (Pevonedistat) was purchased from Cayman Chemicals (Cat. 15217-1). Drugs were added to the medium at the indicated concentrations.

Kim D.K., Redon C.E., Aladjem M.I., Kim H.K, & Jang S.M. (2021). Molecular double clips within RepID WD40 domain control chromatin binding and CRL4-substrate assembly. Biochemical and biophysical research communications, 567, 208-214.

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Myeloma Cell Lines Cultivation and Maintenance

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Three human MM cell lines (LP-1, OPM-2, RPMI-8226) and murine myeloma cell lines (5TGM1, 5TGM1-eGFP, 5T33MMvt) were cultured in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Waltham, Massachusetts, USA) supplemented with 10% fetal bovine serum (FBS) (Biochrom AG, Berlin, Germany), 100 U/mL penicillin, 100 µg/mL streptomycin and 2 mM L-glutamine (Lonza, Basel, Switzerland). The human stromal cell line HS-5 was cultured in DMEM medium supplemented with 10% FBS, 1% sodium pyruvate, 1% MEM NEAA (Thermo Fisher Scientific), 100 U/mL penicillin, 100 µg/mL streptomycin and 2 mM L-glutamine. The human MM cell lines LP-1, OPM-2, RPMI-8226 and HS-5 cells were obtained from ATCC (Molsheim, France) and identity of the cell lines was yearly validated by short-tandem repeat analysis. Cell lines were regularly tested for mycoplasma contamination and passaged no more than 1 month prior to experiments (#LT07-418, Lonza, USA).

Fan R., Satilmis H., Vandewalle N., Verheye E., Vlummens P., Maes A., Muylaert C., De Bruyne E., Menu E., Evans H., Chantry A., De Beule N., Hose D., Törngren M., Eriksson H., Vanderkerken K., Maes K., Breckpot K, & De Veirman K. (2023). Tasquinimod suppresses tumor cell growth and bone resorption by targeting immunosuppressive myeloid cells and inhibiting c-MYC expression in multiple myeloma. Journal for Immunotherapy of Cancer, 11(1), e005319.

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Studying Cell Cycle Regulation

Cited 1 time

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Human U2OS, HCT116, and K562 cells with and without RepID deletion^{37 (link)} were cultured in Dulbecco’s modified Eagle’s medium (Invitrogen, 10569-010) supplemented with 10% heat-inactivated fetal bovine serum in a 37 °C/5% CO₂ humidified incubator. All original cancer cell lines were obtained from ATCC (www.atcc.org). All cell lines were tested for mycoplasmas (Lonza, LT07-418). MLN4924 and SKP2 inhibitors were purchased from Cayman Chemicals (Cat. 15217-1) and Millipore (Cat. 506305), respectively. Drugs were added to the media at the indicated concentrations. HCT116 cells were synchronized in mitosis by a shake-off after 16 h of incubation in 100 nM nocodazole. Mitotic cells were washed three times in phosphate-buffered saline (PBS) and either collected immediately (0 h) or released in fresh medium for different time periods. The cell cycle distribution of cells was confirmed by flow cytometry using LSRFortessa cell analyzer (BD Biosciences) after staining DNA with DAPI.

Jang S.M., Zhang Y., Utani K., Fu H., Redon C.E., Marks A.B., Smith O.K., Redmond C.J., Baris A.M., Tulchinsky D.A, & Aladjem M.I. (2018). The replication initiation determinant protein (RepID) modulates replication by recruiting CUL4 to chromatin. Nature Communications, 9, 2782.

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