Detroit 562

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Detroit 562 is a laboratory instrument designed for high-throughput analysis. It features a compact and modular design, enabling efficient sample processing and data acquisition. The core function of the Detroit 562 is to provide reliable and consistent measurement capabilities for a variety of analytical applications.

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Lab products found in correlation

8 protocols using detroit 562

Genetic Manipulation of Pharyngeal Cancer Cells

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Human pharyngeal carcinoma cells (FaDu and Detroit 562) and human nasopharyngeal epithelial cells (NP69) were obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were cultured in Dulbecco’s modified Eagle medium (Thermo Fisher Scientific, Inc. Waltham, MA, USA) supplemented with 10% fetal calf serum (Invitrogen, Carlsbad, CA, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin (Invitrogen) at 37 °C in a humidified incubator under 5% CO₂.
Short hairpin RNAs (shRNAs) knocking down HOXA11-AS1, PD-L1, and PTBP1 and lentiviral plasmids overexpressing HOXA11-AS1 and PTBP1 were synthesized by RiboBio (Guangzhou, China). All plasmids and their combinations were transduced into FaDu or Detroit 562 cells using Lipofectamine 3000 (Invitrogen).

Zhou Z., Liu Q., Zhang G., Mohammed D., Amadou S., Tan G, & Zhang X. (2022). HOXA11-AS1 Promotes PD-L1-Mediated Immune Escape and Metastasis of Hypopharyngeal Carcinoma by Facilitating PTBP1 and FOSL1 Association. Cancers, 14(15), 3694.

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Modulating cancer cell lines using HOXA11-AS1, PD-L1, and PTBP1

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Human pharyngeal carcinoma cells (FaDu and Detroit 562) and human nasopharyngeal epithelial cells (NP69), were obtained from the American Type Culture Collection (Manassas, VA, USA), and then cultured in Dulbecco's modi ed Eagle medium (Thermo Fisher Scienti c, Inc. Waltham, MA, USA) supplemented with 10% fetal calf serum (Invitrogen, NY, CA, USA), 100U/mL penicillin and 100 μg/mL streptomycin (Invitrogen) at 37˚C in a humidi ed incubator with 5% CO 2 .
The short hairpin RNA (shRNA) knocking down HOXA11-AS1, PD-L1 and PTBP1, the lentiviral plasmids overexpressing HOXA11-AS1 and PTBP1 were synthesized by RiboBio (Guangzhou, China). All plasmids and the combination were transduced into FaDu or Detroit 562 cells using Lipofectamine 3000 (Invitrogen).

Zhou Z., Liu Q., Zhang G., Mohammed D., Amadou S., Tan G., & Zhang X. (2022). HOXA11-AS1 promotes PD-L1 mediated immune escape and metastasis of hypopharyngeal carcinoma by facilitating the association of PTBP1 with FOSL1.

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Cell Line Authentication and Treatment

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The ParC5 (RRID:CVCL_D695) cell line has been previously described (56 (link)). Detroit 562 (ATCC Cat# CCL-138, RRID:CVCL_1171) and FaDu (ATCC Cat# HTB-43, RRID:CVCL_1218) cells were obtained from the CU Anschutz Cell Technologies Shared Resource and cultured in DMEM/High glucose medium (Thermo Scientific, Logan, UT, USA, #SH30243.02) supplemented with 10% FBS (Sigma, St Louis, MO, USA, #F2442) and grown in 5% CO₂ humid cell culture incubator. Cell line profiling for authentication was done through the CU Anschutz Cell Technologies Shared Resource at the University of Colorado Anschutz Medical Campus. Cells used in these experiments were within ten passages of authentication and were monitored for Mycoplasma once a month using the Plasmotest kit from Invivogen (San Diego, CA). For some experiments, subconfluent cells (60–80%) were treated with dasatinib (50 nM) or imatinib (10 μM) (Selleckchem, Houston, TX), or the DMSO vehicle, and irradiated using a Cesium-137 source. Other inhibitors used include NU7441 (DNA-PK), PD98059 (MEK1) (Selleckchem, Houston, TX), Mirin (Rad50/MRN), and B02 (Rad51) (Cayman Chemical, Ann Arbor, MI). The ON-TARGETplus siRNAs (SMARTpool) nontargeting control (siNT, D-001810-10-05) and targeting rat DNA ligase 4 (siLig4, L-089681-02-0005) and rat BRCA1 (siBRCA1, L-103578-02-0005) were purchased from Dharmacon-Horizon Discovery (Lafayette, CO).

Affandi T., Ohm A.M., Gaillard D., Haas A, & Reyland M.E. (2021). Tyrosine kinase inhibitors protect the salivary gland from radiation damage by increasing DNA double-strand break repair. The Journal of Biological Chemistry, 296, 100401.

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In Vitro Evaluation of S. pyogenes Infection

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Human cell lines used in the in vitro studies included preferred niches of colonization/infection of S. pyogenes from the respiratory track: Detroit 562 (ATCC^® CCL138^™), a cell line derived from the metastatic site of pharynx carcinoma, primary Bronchial/Tracheal Epithelial Cells (BTEC) (ATCC^® PCS300010^™), and A549 (ATCC^® CCL185^™), a cell line derived from a human adenocarcinoma of the alveolar basal epithelial cells. All cell lines were purchased from the American Type Culture Collection (ATCC) (www.atcc.org) and cultured according to the manufacturer′s specifications. Detroit 562 and A549 cultures were maintained in Dulbecco's modified eagle medium (DMEM, ThermoFisher Scientific) supplemented with 10% (v/v) Fetal Bovine serum (ThermoFisher Scientific), and a mixture of 100 U/ml penicillin and 100 μg/ml streptomycin (ThermoFisher Scientific). BTEC were maintained in Airway Epithelial Cell Basal medium (ATCC) supplemented with Bronchial epithelial cell growth kit (ATCC), 33 μmol/l Phenol Red (Sigma) and a mixture of 100 U/ml penicillin and 100 μg/ml streptomycin (ThermoFisher Scientific). For bacterial internalization and adherence assays, human cells were seeded in a 96‐well culture plate (Sigma‐Aldrich Co. LLC, St. Louis, United States of America) at a density of 3 × 10⁴ cells/well and incubated for 24 hours at 37°C, 5% (v/v) CO₂ and 99% (v/v) relative humidity.

Alves‐Barroco C., Roma‐Rodrigues C., Raposo L.R., Brás C., Diniz M., Caço J., Costa P.M., Santos‐Sanches I, & Fernandes A.R. (2018). Streptococcus dysgalactiae subsp. dysgalactiae isolated from milk of the bovine udder as emerging pathogens: In vitro and in vivo infection of human cells and zebrafish as biological models. MicrobiologyOpen, 8(1), e00623.

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Cell Culture of Human HNSCC Lines

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Human HNSCC cell lines (Detroit 562, Cal27, SCC-9, SCC-15, and Fadu) and a normal oral epithelial cell line (HOK) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). HEK-293T cell line was obtained from the Shanghai Institute of Life Science, Chinese Academy of Sciences (Shanghai, China). Detroit 562 and Fadu cell lines were grown in the Eagle’s Minimum Essential Medium (EMEM) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Inc., Waltham, MA, USA), penicillin and streptomycin and SCC-9 and SCC-15 cells were cultured in F12/Dulbecco’s Modified Eagle’s Medium (DMEM; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS, penicillin and streptomycin, while Cal27 and HEK-293T cells were maintained in DMEM supplemented with 10% FBS, penicillin and streptomycin. HOK cells were cultured in Oral Keratinocyte Medium (iXCells Biotechnologies, California, USA). All the cell lines were cultured in a humidified incubator with 5% CO₂ at 37 °C.

Huang S., Zhan Z., Li L., Guo H., Yao Y., Feng M., Deng J, & Xiong J. (2019). LINC00958-MYC positive feedback loop modulates resistance of head and neck squamous cell carcinoma cells to chemo- and radiotherapy in vitro. OncoTargets and therapy, 12, 5989-6000.

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Culturing SCC-25 and Detroit 562 Cell Lines

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SCC-25 (CRL1628TM, human papilloma virus [HPV] negative) and Detroit 562 (HPV negative) cells were purchased from ATCC (Manassas, VA, USA). SCC25 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM)/F12 (GibcoBRL, Waltham, MA, USA) supplemented with hydrocortisone (400 ng/mL), 10% heat-inactivated foetal bovine serum (FBS) (Omega Scientific, Inc, Tarzana, CA, USA), 50 IU/mL penicillin and 50 µg/mL streptomycin (GibcoBRL). Detroit 562 cells were cultured in Eagle’s Minimum Essential Medium (EMEM) (GibcoBRL) with 10% FBS, 50 IU/mL penicillin and 50 µg/mL streptomycin (GibcoBRL) in 5% CO₂ at 37 °C. The cells were negative for mycoplasma contamination (MycoAlert Mycoplasma Detection Kit, Lonza, Walkersville, MD, USA).

Perri G., Vilas Boas V.G., Nogueira M.R., Mello Júnior E.J., Coelho A.L., Posadas E.M., Hogaboam C., Cavassani K.A, & Campanelli A.P. (2024). Interleukin 33 supports squamous cell carcinoma growth via a dual effect on tumour proliferation, migration and invasion, and T cell activation. Cancer Immunology, Immunotherapy : CII, 73(6), 110.

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Cell Viability Assay for Pharyngeal Cells

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For the cell viability experiment, the human pharyngeal cell line Detroit 562
(ATCC CCL-138) was purchased from American Type Culture Collection (Manassas,
VA). The Detroit 562 cell line was cultured in Modified Eagle’s Medium
(Gibco-BRL, Grand Island, NY) supplemented with 10% fetal bovine serum
(Gibco-BRL, Grand Island, NY), 100 U/mL penicillin, and 100 µg/mL streptomycin
at 37°C in a humidified atmosphere of 5% CO₂.
The cell proliferation experiment was performed using the
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. For 24
hours, the Detroit 562 cells were starved and simultaneously treated with each
concentration of OCE (1, 5, 10, 50, or 100 µg/mL) or with each concentration of
OCE plus 10 µM 5-FU. After 48 hours, the medium was removed and the cells were
incubated with MTT to measure metabolic activity. To measure metabolic activity,
spectrophotometric analysis at 450 nm was performed using a microtiter plate
reader (Molecular Devices, LLC, Sunnyvale, CA).

Park J.W., Oh J., Ko S.J., Chang M.S, & Kim J. (2018). Effects of Onchung-eum, an Herbal Prescription, on 5-Fluorouracil–Induced Oral Mucositis. Integrative Cancer Therapies, 17(4), 1285-1296.

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Establishing In Vitro Models for Cancer Research

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Human head and neck cancer cell lines (CAL27 (CRL2095), Detroit 562 (CCL-138)), murine lung (LL/2 (CRL-1642)) and murine melanoma (B16 (CRL-6475)) were obtained from American Type Culture Collection. Murine colorectal (MC38 (ENH204-FP)) cancer cell lines were obtained from Kerafast. Murine head and neck MOC1 and MOC2 were generously provided by Dr. R. Uppaluri. CAL27 PIK3CA H1047R expressing cells and 4MOSC1 cells were generated in Dr. Gutkind’s laboratory and previously characterized^{42 (link),55 (link)}. CAL27, Detroit 562, LL/2, and B16 cells were cultured in DMEM (Gibco) supplemented with 10% FBS (Omega Scientific). MC38 cells were cultured in DMEM supplemented with 10% FBS, 1 mM sodium pyruvate (Gibco), 1% non-essential amino acids (Gibco), and 10 mM HEPES (Gibco). MOC1 and MOC2 cells were cultured in IMDM (Lonza)/F-12K nutrient mixture at a 2:1 mixture with 5% FBS, 5 ng/ml EGF (EMD Millipore), 400 ng/ml hydrocortisone (Sigma), and 5 mg/ml insulin (Sigma). On receipt, each cell line was expanded, cryopreserved as low passage stocks, and routinely tested for mycoplasma. Cisplatin (Enzo Biosciences), paclitaxel (Sigma), and MMAE (Concortis) were reconstituted in DMSO. Clinical-grade CDX3379 was provided by Celldex. Clinical grade cetuximab (Erbitux, Lilly Medical) was obtained from UCSD Moores Cancer Center Pharmacy.

Hingorani D.V., Allevato M.M., Camargo M.F., Lesperance J., Quraishi M.A., Aguilera J., Franiak-Pietryga I., Scanderbeg D.J., Wang Z., Molinolo A.A., Alvarado D., Sharabi A.B., Bui J.D., Cohen E.E., Adams S.R., Gutkind J.S, & Advani S.J. (2022). Monomethyl auristatin antibody and peptide drug conjugates for trimodal cancer chemo-radio-immunotherapy. Nature Communications, 13, 3869.

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