Rt112

Manufactured by Thermo Fisher Scientific

Sourced in United States

The RT112 is a real-time PCR thermal cycler designed for DNA amplification and quantification. It features precise temperature control and rapid ramp rates to ensure accurate and reliable results. The RT112 supports a wide range of sample volumes and can accommodate multiple plate formats.

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Lab products found in correlation

9 protocols using rt112

Conditional Expression of p15/p16 in Bladder Cancer

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Two human bladder cancer cell lines, RT112 and UMUC3, that had 9p21.3 deletion^{37 (link)} were purchased from the American Type Culture Collection (Manassas, VA, USA) and used for all transfection experiments. Both the cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Grand Island, NY, USA) supplemented with 10% Tet-system approved fetal bovine serum (Clontech, Mountain View, CA, USA) at 37 °C with 5% CO₂.
A tetracycline-based inducible system was chosen for conditional gene expression. Briefly, plasmid pTet-on (Clontech, Mountain View, CA, USA) was transfected into RT112 and UMUC3 cells using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA), followed by selection of stable clones in 300 μg/ml G418 (Gibco, Grand Island, NY, USA). pTRE2-pur containing p15, p16, p15-N/p16-C or p16-N/p15-C was then transfected into pTet-on-stably transfected RT112 or UMUC3 using Lipofectamine 2000 and stable clones were selected in the presence of puromycin (Gibco) (0.8 μg/ml for RT112 and 1.2 μg/ml for UMUC3).

Xia Y., Liu Y., Yang C., Simeone D.M., Sun T.T., DeGraff D.J., Tang M.S., Zhang Y, & Wu X.R. (2021). Dominant role of CDKN2B/p15INK4B of 9p21.3 tumor suppressor hub in inhibition of cell-cycle and glycolysis. Nature Communications, 12, 2047.

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Bladder Cancer Cell Lines Cultivation

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The human urinary bladder transitional cell carcinoma cell lines T24 (RRID: CVCL_0554), UM-UC-3 (RRID: CVCL_1783) were purchased from ATCC, RT112 (RRID: CVCL_1670) was purchased from German Collection of Microorganisms and Cell Cultures GmbH (DSMZ) and UM-UC-1 (RRID: CVCL_2743) was purchased from European Collection of Authenticated Cell Cultures (ECACC). Human normal bladder epithelial cell line SV-HUC-1 (RRID: CVCL_3798) was purchased from ATCC. Murine transitional cell carcinoma cell line MB49 (RRID: CVCL_7076) was purchased from Millipore, and murine bladder epithelial cells (MBEC) were purchased from Procell (catalog no. CP-M058). Human lymphatic endothelial cells (HLEC) were obtained from ScienCell Research Laboratories. All cells were maintained in a humidified incubator with 5% CO₂ at 37°C. The T24, RT112, and UM-UC-1 cells were cultured in RPMI 1640 medium (Gibco, catalog no. C11875500BT). The UM-UC-3 and MB49 cells were cultured in DMEM (Gibco, catalog no. C11995500BT). The SV-HUC-1 cells were cultured in Ham's F12K medium (Gibco, catalog no. 21127022). All medium was supplemented with 10% FBS (BI, catalog no. 04–001–1ACS). The HLECs were cultured in endothelial cell medium (ECM) with 5% FBS (ScienCell Research Laboratories, catalog no. 1001). The authentication and Mycoplasma testing of all cell lines were qualified.

An M., Zheng H., Huang J., Lin Y., Luo Y., Kong Y., Pang M., Zhang D., Yang J., Chen J., Li Y., Chen C, & Lin T. (2022). Aberrant Nuclear Export of circNCOR1 Underlies SMAD7-Mediated Lymph Node Metastasis of Bladder Cancer. Cancer Research, 82(12), 2239-2253.

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Bladder Cancer Cell Line Transfection

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Human ureteral epithelial cells SV-HUC-1, BC cell lines including UM-UC-1, UM-UC-3, RT4, RT112 and T24 were purchased from Procell Life Sciences (Wuhan, China). SV-HUC-1 cells were cultured in Ham’s F-12 K medium (Macgene, China) supplemented with 10% fetal bovine serum (FBS) (Gibco, MA, USA). UM-UC-1, UM-UC-3, RT4, RT112 and T24 cells were cultured in RPMI-1640 medium (Gibco, MA, USA) supplemented with 10% FBS. All cells were cultured in 5% CO₂ at 37 °C.
The si-PFK-1 and PFK-1 overexppressing plasmids and their negative control were synthesized by Gemma Shanghai. UM-UC-1 and RT112 cells were grown at 90% fusion. Transfection was performed using Lipofectamine 3000 regent according to the instructions. The culture medium was changed 6 h after transfection, and PCR was performed to detect transfection efficiency at 24–48 h after transfection.

Wang R., Xu F., Yang Z., Cao J., Hu L, & She Y. (2024). The mechanism of PFK-1 in the occurrence and development of bladder cancer by regulating ZEB1 lactylation. BMC Urology, 24, 59.

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Cell Culture Protocols for Bladder Cancer Lines

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The cell line RT112 (DSMZ) was maintained in MEM alpha medium with GlutaMAX supplemented with 10% FBS (fetal bovine serum, both Gibco, Life Technologies, Waltham, USA). The cell line and their knockout derivatives were validated to be RT112 by single nucleotide polymorphism (SNP) profiling (performed by Multiplexion GmbH, Friedrichshafen, Germany). Cell line J82 (ATCC) and HS853T (ATCC) were cultured in DMEM medium (4.5 g/L glucose) supplemented with 10% FBS (both Gibco, Life Technologies, Waltham, USA), 1% HEPES solution, and 1% MEM non-essential amino acids (both Sigma-Aldrich, St. Louis, USA). The cell line 5637 (ATCC) was cultured in RPMI supplemented with 10% FBS (both Gibco, Life Technologies, Waltham, USA). All cell lines were subject to regular testing for excluding mycoplasma contamination (last on 2nd April 2019). All cells were maintained at standard conditions in a humidified incubator with 5% CO₂ at 37°C.

Schulz A., Gorodetska I., Behrendt R., Fuessel S., Erdmann K., Foerster S., Datta K., Mayr T., Dubrovska A, & Muders M.H. (2020). Linking NRP2 With EMT and Chemoradioresistance in Bladder Cancer. Frontiers in Oncology, 9, 1461.

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Bladder Cancer Cell Line Culture

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The SV-HUC-1 (CL-0222), HT-1376 (CL-0672), BIU-87 (CL-0035), T24 (CL-0227), RT4 (CL-0431) and 5,637 (CL-0002) cell lines were purchased from Procell Life Science and Technology Co., Ltd., Wuhan, China. The RT-112 (C6779) cell was purchased from Beyotime. SV-HUC-1 cells were cultured in Ham’s F-12K medium supplemented with 10% foetal bovine serum (FBS, Gibco); HT-1376 cells were cultured in MEM supplemented with 10% FBS; BIU-87, T24, RT4, RT-112 and 5,637 cells were cultured in RPMI-1640 supplemented with 10% FBS.

Liu J., Zhang Z., Zhang W., Meng L., Wang J., Lv Z., Xia H., Wu M., Zhang Y, & Wang J. (2022). Ferroptosis Mediation Patterns Reveal Novel Tool to Implicate Immunotherapy and Multi-Omics Characteristics in Bladder Cancer. Frontiers in Cell and Developmental Biology, 10, 791630.

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Culturing Bladder Cell Lines

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We obtained three bladder cell lines (RT-112, T24, and 5637) and a normal human bladder epithelial cell line (SV-HUC-1) from BeNa Culture Collection (BNCC, China). RT-112 and 5637 cell lines were cultured in RPMI-1640 medium (Gibco, USA), T24 cell line in McCoy's 5a medium (Gibco, USA), and SV-HUC-1 cell line in F-12K medium. All the cells were cultured with 10% FBS (Gibco, USA) and the condition of 5% CO₂ and 37°C.

Jiang J., Zhan Y, & Li J. (2022). The Role of the Key Differentially Mutated Gene FGFR3 in the Immune Microenvironment of Bladder Cancer. Journal of Immunology Research, 2022, 7952706.

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Cell Line Cultivation Conditions

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RT-112 and VMCUB-1 cells were obtained from Leibniz Institut DSMZ, Germany. HT-1376, 5637, HT-1197 and T24 were purchased from ATCC. RT-112, 5637, VMCUB-1 cells were cultivated at 37 °C in a humidified environment with 5% CO² in RPMI medium (Gibco™ RPMI 1640 Medium, Life Technologies™), while HT-1376 and HT-1197 were cultivated in MEM medium (Gibco MEM (1x), Life Technologies™) and T24 in McCoy's 5a modified medium (Gibco™ McCoy's 5A (Modified) Medium, Life Technologies™). All media was supplemented with 10% fetal bovine serum and 100 U/ml penicillin G and 100 μg/ml streptomycin.

Eich M.L., Rodriguez Pena M.D., Chandrashekar D.S., Chaux A., Agarwal S., Gordetsky J.B., Ferguson J.E., Sonpavde G.P., Netto G.J, & Varambally S. (2019). Expression and Role of Methylenetetrahydrofolate Dehydrogenase 1 Like (MTHFD1L) in Bladder Cancer. Translational Oncology, 12(11), 1416-1424.

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Bladder Cancer Cell Line Cultures

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T24, J82, HT1376, 5637, RT4, and RT112 bladder cancer cell lines were used in this study. J82 and RT4 cell lines were kindly provided by Şerif Şentürk and T24 cell line was provided by Neşe Atabey. Other cell lines HT1376, RT112, and 5637 were purchased from DSMZ (German Collection of Microorganisms and Cell Lines). T24, J82, RT4, and HT1376 cell lines were grown with Dulbecco’s modified Eagle medium (Gibco) supplemented with 10% fetal bovine serum and 1% penicillin. Other cell lines, RT112 and 5637 were grown in Roswell Park Memorial Institute (RPMI) 1640 Medium (Gibco) supplemented with 10% fetal bovine serum and 1% penicillin. These cell lines were tested (EZ-PCR Mycoplasma Detection Kit, Cat. No:20-700-20) for mycoplasma contamination and the results were negative. C2C12 myoblast cells were kindly provided by Mehmet Öztürk. Muscle differentiation of C2C12 cells was achieved using 10% horse serum (Gibco, 26050070) for 5 days.

Guneri-Sozeri P.Y., Özden-Yılmaz G., Kisim A., Cakiroglu E., Eray A., Uzuner H., Karakülah G., Pesen-Okvur D., Senturk S, & Erkek-Ozhan S. (2023). FLI1 and FRA1 transcription factors drive the transcriptional regulatory networks characterizing muscle invasive bladder cancer. Communications Biology, 6, 199.

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Establishing RCC Cell Lines for Research

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Thirty pairs of RCC specimens and adjacent normal tissues were obtained from patients who underwent surgical treatment from October 2015 to August 2016 in Tai'an City Central Hospital. Written informed consent was obtained prior to resection from patients and the study was approved by the Ethics Committee of Tai'an City Central Hospital.
Cell lines. RCC cell lines 786-O, Caki-1, Ketr-3, RT112 and T24, as well as the normal renal epithelial cells Ect1/E6E7 were purchased from the American Type Culture Collection (ATCC, Gaithersburg MD, USA). The 786-O, Ketr-3, RT-112 and T24 cells were cultured in RPMI-1640 medium (Gibco, Rockville, MD, USA) containing 10% fetal bovine serum (FBS; Gibco). The Caki-1 cells were cultured in McCoy's 5A medium containing 10% FBS, and Ect1/E6E7 cells were cultured in EMEM medium (Gibco) containing 10% FBS. All cells were cultured in incubators at 37˚C in a 5% CO 2 atmosphere.

Li Z., Ma Z, & Xu X. (2019). Long non‑coding RNA MALAT1 correlates with cell viability and mobility by targeting miR‑22‑3p in renal cell carcinoma via the PI3K/Akt pathway. Oncology reports, 41(2).

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