Ll 2 luc m38

Manufactured by PerkinElmer

Sourced in United States

The LL/2-luc-M38 is a cell line that has been genetically modified to express the luciferase reporter gene. This cell line can be used in various in vitro assays that involve bioluminescence detection.

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Lab products found in correlation

Dmem with 4.5 g l glucose, by Corning (1 mentions) Rpmi 1640, by Corning (1 mentions) Dmem medium, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) A549luc, by PerkinElmer (1 mentions) Mda mb 231 luc, by PerkinElmer (1 mentions) Ht 29 luc, by PerkinElmer (1 mentions)

5 protocols using ll 2 luc m38

Generating Stable Luciferase-Expressing Cell Lines

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CMT167 cells (12 (link)) were stably transfected with firefly luciferase as previously described (11 (link)). Lewis Lung Carcinoma Cells (LL/2) were purchased from ATCC and luciferase-expressing Lewis Lung Carcinoma cells (LLC) were purchased from Caliper Life Sciences (LL/2-luc-M38). All cell lines were periodically tested for mycoplasma infection and were last retested in February 2017. To avoid cross-contamination and phenotypic changes, cells were maintained as frozen stocks and cultured for only two to four weeks before use in experiments. Authentication of cell lines based on morphology, growth curve analysis, and metastatic phenotype was performed regularly, and no phenotypic changes were observed through the duration of the study.

Li H.Y., McSharry M., Bullock B., Nguyen T.T., Kwak J., Poczobutt J.M., Sippel T.R., Heasley L.E., Weiser-Evans M.C., Clambey E.T, & Nemenoff R.A. (2017). The Tumor Microenvironment Regulates Sensitivity of Murine Lung Tumors to PD-1/PD-L1 Antibody Blockade. Cancer immunology research, 5(9), 767-777.

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Stable Luciferase-Expressing KRAS Mutant Cell Lines

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CMT167 cells were stably transfected with firefly luciferase, as previously described (24 (link)). Luciferase-expressing LLC cells were purchased from Caliper Life Sciences (LL/2-luc-M38). The CMT167 cells harbor a Kras^G12V mutation, and the LLC cells harbor a Kras^G12C mutation (24 (link)). Both cell lines were maintained in DMEM with 4.5 g/l glucose (no. 10–017-CV; Corning) containing 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 500 μg/ml G418 at 37°C in a humidified 5% CO₂ atmosphere. Cells were periodically tested for mycoplasma infection, maintained as frozen stocks, and cultured for only 2–4 wk before use in experiments. Authentication of cell lines based on morphology, growth curve analysis, and metastatic phenotype was performed regularly. Human NSCLC cell lines were maintained in RPMI 1640 (no. 10–040-CV; Corning) containing 10% FBS at 37°C in a humidified 5% CO₂ atmosphere. All were KRAS mutant.

Johnson A.M., Bullock B.L., Neuwelt A.J., Poczobutt J.M., Kaspar R.E., Li H.Y., Kwak J.W., Hopp K., Weiser-Evans M.C., Heasley L.E., Schenk E.L., Clambey E.T, & Nemenoff R.A. (2020). Cancer Cell–Intrinsic Expression of MHC Class II Regulates the Immune Microenvironment and Response to Anti–PD-1 Therapy in Lung Adenocarcinoma. Journal of immunology (Baltimore, Md. : 1950), 204(8), 2295-2307.

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Stable Luciferase-Expressing Cell Lines

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CMT167 cells (13 (link)), originally provided by Dr. Alvin Malkinson (University of Colorado), were stably transfected with firefly luciferase as previously described (12 (link)) and maintained in DMEM with 1 g/L glucose containing 10% FBS, 100 U/mL penicillin, 100 μg/ml streptomycin, MEM Non-Essential Amino Acids, 10 mM HEPES, and 500 μg/mL G418 at 37°C in a humidified 5% CO₂ atmosphere. Luciferase-expressing Lewis Lung Carcinoma (LLC) cells were purchased from Caliper Life Sciences (LL/2-luc-M38) and maintained in DMEM with 4.5 g/L glucose containing 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, and 500 μg/mL G418 at 37°C in a humidified 5% CO₂ atmosphere. Both cell lines were periodically tested for mycoplasma infection and were last retested in October 2016. To avoid cross-contamination and phenotypic changes, these cell lines have not been maintained in long-term cultures. All cells used in this study were maintained as frozen stocks and cultured for only 2–4 weeks before use in experiments. Authentication of these cell lines based on morphology, growth curve analysis, and metastatic phenotype was performed regularly, and no phenotypic changes were observed through the duration of the study.

Kwak J.W., Laskowski J., Li H.Y., McSharry M.V., Sippel T.R., Bullock B.L., Johnson A.M., Poczobutt J.M., Neuwelt A.J., Malkoski S.P., Weiser-Evans M.C., Lambris J., Clambey E.T., Thurman J.M, & Nemenoff R.A. (2017). Complement activation via a C3a receptor pathway alters CD4+ T lymphocytes and mediates lung cancer progression. Cancer research, 78(1), 143-156.

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NSCLC Cell Line Establishment and Validation

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NSCLC cell lines, including A549, Calu-3, SK-MES-1, PAa, NCI-H292, NCI-H520 (H520), NCI-H596, NCI-H1299, NCI-H1650, NCI-H1975, HLAMP and 95D were obtained from cell banks of Shanghai Institutes of Biological Sciences (Shanghai, China) or Fu Erbo Biotechnology Co., Ltd. (Guangzhou, China). Murine lung carcinoma cell line LL/2-luc-M38 was provided by Caliper Life Sciences, Inc (Massachusetts, USA). Subcutaneous tumours-derived cells were primarily cultured and primary normal lung epithelial cells were obtained instructed by the protocol of previous report51 (link). NSCLC cell lines were maintained in DMEM medium (Invitrogen) supplemented with 10% fetal bovine serum (HyClone, Logan, UT) and 1% penicillin/streptomycin (Invitrogen). The BEAS-2B immortalized human bronchial epithelial cell line (Shanghai Institutes of Biological Sciences, Shanghai, China) was cultured in LHC-9 medium as instructed by the provider. All cell lines were
authenticated by short tandem repeat fingerprinting at IDEXX RADIL (Columbia, MO, USA) and Services at SYSU Forensic Medicine Lab (Guangzhou, China), and were free of mycoplasma contamination.

Cai J., Fang L., Huang Y., Li R., Xu X., Hu Z., Zhang L., Yang Y., Zhu X., Zhang H., Wu J., Huang Y., Li J., Zeng M., Song E., He Y., Zhang L, & Li M. (2017). Simultaneous overactivation of Wnt/β-catenin and TGFβ signalling by miR-128-3p confers chemoresistance-associated metastasis in NSCLC. Nature Communications, 8, 15870.

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Primary Tumor Cell Isolation Protocol

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A549-luc, BXPC-3-luc, HT-29-luc, Hela-luc, MDA-MB231-luc, NCIH460-luc, PC-3M-luc, and LL/2-luc-M38 cell lines were all purchased from Caliper Life Sciences. U87MG-luc cells were generated by our group previously [35 ]. The primary culture was derived from freshly isolated tumor cell from patients of two primary lung cancer or two primary breast cancer individually, in Beijing Shijitan Hospital of Capital Medical University. The consent was obtained from the patients before sample collection. The study complied with the Declaration of Helsinki and was approved by the Biomedical Research Ethics Committee of Beijing Shijitan Hospital of Capital Medical University. Briefly, tumors were placed into RPMI1640 medium with antibiotics and anti-fungal immediately following resection. Specimens were washed with cold PBS repeatedly to remove blood clots and tissue debris, and then minced into 0.5- to 1.0-mm pieces. The tissue pieces were digested with collagenase IV, Dnase I and hyaluronidase. Then cells were washed and passed through a 70 μm cell strainer to obtain single-cell population. All agents for cell culture were from Gibco Company (Gaithersburg, MD, USA). Beige-SCID Mice (8 to 10 weeks) were purchased from Vital River Laboratories (VRL, Beijing, China).

Ma J., Ma P., Zhao C., Xue X., Han H., Liu C., Tao H., Xiu W., Cai J, & Zhang M. (2016). B7-H3 as a promising target for cytotoxicity T cell in human cancer therapy. Oncotarget, 7(20), 29480-29491.

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