Granta 519

Manufactured by Thermo Fisher Scientific

Sourced in United States

The GRANTA-519 is a laboratory equipment designed for thermal analysis. It is capable of measuring the thermal properties of materials, such as their melting point, glass transition temperature, and specific heat capacity. The GRANTA-519 utilizes advanced thermal analysis techniques to provide accurate and reliable data on the thermal behavior of samples.

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6 protocols using granta 519

Targeting Epigenetic Regulators in MCL

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Four MCL cell lines Granta-519, JVM-2, Mino, and Z138 were purchased from the ATCC (Manassas, VA, USA). Granta-519 was cultured with Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, USA) and the rests were cultured with RPMI-1640 medium (Life Technologies, California, USA). Both media were supplemented with 10% fetal bovine serum (FBS) (Gibco, USA) and 1% penicillin/streptomycin. HEK293T cells were routinely maintained in the lab and grown in DMEM supplemented with 10% FBS. All cells were incubated at 37°C in a humidified atmosphere containing 5% CO₂. Cells were plated at a density of 0.1 × 10⁶ cells/flask in T25 flask for cell viability assay, at 0.5 × 10⁶ cells/flask for apoptosis assays and cell cycle analysis, and at 1 × 10⁶ cells/flask for Western blot. EPZ005687 and UNC1999 were purchased from Selleckchem (Houston, TX, USA). The compound was dissolved in anhydrous dimethyl sulfoxide (DMSO) to a stock solution of 10 mM, and then further diluted to the desired concentration for in vitro experiments.

Li W., Bi C., Han Y., Tian T., Wang X., Bao H., Xu X., Zhang X., Liu L., Zhang W., Gao H., Wang H., Zhang H., Meng B., Wang X, & Fu K. (2019). Targeting EZH1/2 induces cell cycle arrest and inhibits cell proliferation through reactivation of p57CDKN1C and TP53INP1 in mantle cell lymphoma. Cancer Biology & Medicine, 16(3), 530-541.

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Characterization of MCL cell lines

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We used two well characterized MCL cell lines (13 (link)), JeKo-1 (ATCC, CRL-3006, RRID:CVCL_1865) and Granta-519 (DSMZ, ACC-342, RRID:CVCL_1818), the lymphoblastoid leukemic cell line JVM13 (ATCC, CRL-3003, RRID:CVCL_1318), and HEK293T (ATCC, CRL-3216, RRID:CVCL_0063). Cell lines were cultured at 37°C and 5% CO₂; JeKo-1 and JVM13 in RPMI (Gibco) and Granta-519 and HEK293T in DMEM (Gibco), both supplemented with 10% FBS and penicillin-streptomycin (Gibco). Cell line authentication was performed by qCell Identity (qGenomics) when appropriate. Cells were tested for Mycoplasma on a regular basis. We obtained cyclin D1 knocked-down MCL cell lines by lentiviral infection using the shRNA plasmids TRCN0000295873 and TRCN0000295874 (Sigma-Aldrich) as described before (20 (link),21 (link)). Cyclin D1 inducible overexpression models in JVM13, with wild type (CycD1^wt) or a highly stable form of the protein with the T286A mutation (CycD1^T286A), were generated previously (20 (link)). Cell infections and gene expression analyses were performed with low-passage cells.

Demajo S., Albero R., Clot G., Castellano G., Navarro A., Capdevila C., Enjuanes A., Nadeu F., Giné E., Pinyol M., Jaffe E.S., Ott G., Staudt L.M., Rosenwald A., Scott D.W., Rimsza L.M., López-Guillermo A., Beà S., Campo E, & Jares P. (2020). A cyclin D1-dependent transcriptional program predicts clinical outcome in mantle cell lymphoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(1), 213-225.

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Maintenance of Human MCL Cell Lines

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Human MCL cell lines (REC-1, Z-138, JEKO-1, MINO, GRANTA-519 and UPN-1) were grown in Advanced-RMPI 1640 supplemented with 5% heat-inactivated fetal bovine serum (FBS), 2 mmol/L glutamine, and 50 µg/mL penicillin-streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). JEKO-1, MINO, and REC-1 parental cells were obtained from ATCC cell bank (LGC Standards); GRANTA-519 cell line was purchased at DSMZ; and UPN-1 and Z-138 cells were provided by Dr. B. Sola (University of Caen, Caen, France). Cultures were routinely subjected to cell authentication and mycoplasma detection tests.

Santos J.C., Profitós-Pelejà N., Ribeiro M.L, & Roué G. (2022). Antitumor Activity of Simvastatin in Preclinical Models of Mantle Cell Lymphoma. Cancers, 14(22), 5601.

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Cell Line Culturing Protocol

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The human cell lines ALL-SIL, SKW-3/KE-37, CTV-1, MEC-1, JEKO-1, REC-1, and Granta-519 were purchased from the Leibniz Institut DSMZ-German collection of microorganisms and cell cultures (Germany). Cells were cultured in RPMI 1640 (#MT10040CV, Thermo Fisher Scientific, Waltham MA, USA) with 10% or 20% fetal bovine serum (FBS) (#10270–106, Thermo Fisher Scientific), 1% penicillin-streptomycin (P/S) (#3MT30002CI, Thermo Fisher Scientific), and 1% of MEM Non-Essential Amino Acids Solution (100X) (#11140050, Thermo Fisher Scientific), 2 mM L-Glutamine (#25030-081, Thermo Fisher Scientific), and 1% HEPES Buffer 1M (#MS013D1006, Biowest, Nuaillé, France). Granta-519 was maintained in DMEM (#11960-044, Thermo Fisher Scientific) with 20% FBS, 1% P/S, and 2 mM L-Glutamine (#25030-081, Thermo Fisher Scientific). Cell lines were grown in a humidified incubator at 37 °C, and 5% CO₂ and monitored for mycoplasma contamination.

Pagliaro L., Cerretani E., Vento F., Montanaro A., Moron Dalla Tor L., Simoncini E., Giaimo M., Gherli A., Zamponi R., Tartaglione I., Lorusso B., Scita M., Russo F., Sammarelli G., Todaro G., Silini E.M., Rigolin G.M., Quaini F., Cuneo A, & Roti G. (2024). CAD204520 Targets NOTCH1 PEST Domain Mutations in Lymphoproliferative Disorders. International Journal of Molecular Sciences, 25(2), 766.

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Culturing MCL Cell Lines

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The MCL cell lines JEKO-1 and MINO (American Type Culture Collection) were maintained in RPMI1640 supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin (100 units/ml and 100 g/ml, respectively) (all from Thermo Fisher Scientific). The cell line GRANTA-519 was grown in medium containing 90% minimum essential media and 10% FBS, supplemented with penicillin/streptomycin (cell line and media from Thermo Fisher Scientific). All cell lines were obtained from the American Type Culture Collection and cultured at 37 °C in a humidified, 5% carbon dioxide atmosphere, and tested for mycoplasma contamination before experiments.

Liu H., Lu Z., Shi X., Liu L., Zhang P., Golemis E.A, & Tu Z. (2021). HSP90 inhibition downregulates DNA replication and repair genes via E2F1 repression. The Journal of Biological Chemistry, 297(2), 100996.

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Culturing Diverse Hematological Cell Lines

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All human cell lines (MOLM13, MV4-11, NB4, HL60, OCI-AML3, JVM3, and Granta519) were grown in RPMI1640 media (Thermo Fisher Scientific) supplemented with 10% FBS, GlutaMAX, and 1% Penicillin/Streptomycin (Gibco). MOLM13, NB4, OCI-AML3, JVM3, and Granta519 cells were obtained from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures. MV4-11, HL60, and TF-1 cells were obtained from the ATCC.

Han Y.C., Kahler J., Piché-Nicholas N., Hu W., Thibault S., Jiang F., Leal M., Katragadda M., Maderna A., Dushin R., Prashad N., Charati M.B., Clark T., Tumey L.N., Tan X., Giannakou A., Rosfjord E., Gerber H.P., Tchistiakova L., Loganzo F., O'Donnell C.J, & Sapra P. (2021). Development of Highly Optimized Antibody-Drug Conjugates against CD33 and CD123 for Acute Myeloid Leukemia. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(2).

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