Atcc crl 3216

Manufactured by American Type Culture Collection

Sourced in United States

The ATCC CRL-3216 is a cell line derived from human embryonic kidney cells. It is used for a variety of applications in cell biology and biotechnology research.

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Lab products found in correlation

Hek293t, by American Type Culture Collection (3 mentions) Atcc ccl 10, by American Type Culture Collection (1 mentions) Bhk 21, by American Type Culture Collection (1 mentions) Sk mel 2, by American Type Culture Collection (1 mentions) Colo679, by American Type Culture Collection (1 mentions) Rpmi 7951, by American Type Culture Collection (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Pcdna3, by Thermo Fisher Scientific (1 mentions) Deae dextran, by Merck Group (1 mentions) Fugene 6, by Promega (1 mentions) Lenti x concentrator, by Takara Bio (1 mentions) Sodium pyruvate, by GE Healthcare (1 mentions) Hek293t, by Thermo Fisher Scientific (1 mentions) Non essential amino acids (neaa), by GE Healthcare (1 mentions) Heparin, by Harvard Bioscience (1 mentions) Hepg2, by Thermo Fisher Scientific (1 mentions)

11 protocols using atcc crl 3216

Ethical Animal Handling for Macaque Research

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All animals were handled in strict accordance with good animal practice as defined by the European directive 63/2010/EU and in accordance with recommendations of the Weatherall report. Four to six-year-old cynomolgus macaques (Macaca fascicularis) were imported from the international accredited breeding facilities from Mauritius (negative for SIV, STLV, herpes B virus, filoviruses, SRV-1, SRV-2, measles, dengue virus and CHIKV) and were housed in a BSL3 facility (Permit Number A 92-032-02), in accordance with Office for Laboratory Animal Welfare (OLAW, USA; #A5826-01) standards at the CEA in accordance with the French national regulation under the number B-92-032-02 for animal use, under the number 2005-69 for macaque breeding. Animals are fed daily and monitored closely by caretakers reporting directly to the veterinarians in charge of the animal facilities. All studies were reviewed and approved by the regional animal care and use committee in accordance with European directive 63/2010/EU: “Comité regional d’éthique pour l’expérimentation animale Ile-de-France Sud”, Fontenay aux Roses, decision #07_012.
Cell lines originally purchased from American Tissue Culture Collection (ATCC), such as human embryonic kidney (HEK 293T, ATCC CRL-3216) and baby hamster kidney (BHK21, ATCC CCL-10) cells, were adhered to recommended ethics approvals and standards.

Kam Y.W., Lee W.W., Simarmata D., Le Grand R., Tolou H., Merits A., Roques P, & Ng L.F. (2014). Unique Epitopes Recognized by Antibodies Induced in Chikungunya Virus-Infected Non-Human Primates: Implications for the Study of Immunopathology and Vaccine Development. PLoS ONE, 9(4), e95647.

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Culturing 293T Cells in DMEM

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293T cells were purchased from ATCC (ATCC® CRL-3216) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) with 10% Fetal Bovine Serum.

Yang B.H., Wang K., Wan S., Liang Y., Yuan X., Dong Y., Cho S., Xu W., Jepsen K., Feng G.S., Lu L.F., Xue H.H, & Fu W. (2019). TCF1 and LEF1 Control Treg Competitive Survival and Tfr Development to Prevent Autoimmune Diseases. Cell reports, 27(12), 3629-3645.e6.

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Cell Line Culture Conditions

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293 T, RPMI7951, CJM, A375, SKMEL2, COLO679, and G361 cell lines were purchased from commercial sources (ATCC CRL-3216, ATCC HTB-66, Creative Bioarray CSC-C6421J, ATCC CRL-1619, ATCC HTB-68, Sigma 87061210, and ATCC CRL-1424, respectively) and cultured in DMEM (Life Technologies), except for G361 (McCoy’s media, 10% FBS (Atlanta Biologicals), 1X GlutaMAX (Life Technologies) and 1% penicillin-streptomycin (Life Technologies)). Cells were grown at 37 °C in 5% CO₂. All cultures were regularly checked for the presence of mycoplasma. For cell culture experiments implicating genetic or pharmacological modifications, cells were randomly distributed between the different experimental conditions.

Ablain J., Al Mahi A., Rothschild H., Prasad M., Aires S., Yang S., Dokukin M.E., Xu S., Dang M., Sokolov I., Lian C.G, & Zon L.I. (2022). Loss of NECTIN1 triggers melanoma dissemination upon local IGF1 depletion. Nature Genetics, 54(12), 1839-1852.

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Culturing 293T Cells in DMEM

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293T cells were purchased from ATCC (ATCC® CRL-3216) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) with 10% Fetal Bovine Serum.

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Panx2 Expression and Characterization

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Media, supplements and reagents were obtained from GIBCO^® and Invitrogen™ (Carlsbad, CA, USA). Normal rat kidney (NRK) (ATCC^® CRL-6509™) and human embryonic kidney cells (HEK293T) (ATCC^® CRL-3216™) were obtained from ATCC (Manassas, VA, USA). Adherent HEK293 cells (AD293, Cat# 240085) were obtained from Agilent Technologies, Inc. (Santa Clara, CA, USA). Cell cultures were grown in high-glucose DMEM supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, 100 µg/mL streptomycin and 2mM L-Glutamine. At ~50% of confluency, cells were transfected adding Lipofectamine 3000 (Invitrogen™) following manufacturer directions. 2 µg of pcDNA3.1 (Invitrogen™) plasmids encoding mouse Panx2 [22 (link)] or Panx2^N86Q or their respective FLAG-tagged versions were used for transfections in 35 mm culture plates. After 48 h for single transfections and 72 h for co-transfections, proteins were extracted, and expression levels were determined by Western blot. For co-transfections experiments with mPanx1 plasmid [11 (link)], levels were reduced to 0.5 µg of DNA.

Sanchez-Pupo R.E., Johnston D, & Penuela S. (2018). N-Glycosylation Regulates Pannexin 2 Localization but Is Not Required for Interacting with Pannexin 1. International Journal of Molecular Sciences, 19(7), 1837.

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Culturing 293T Cells with FBS

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The 293T cells (ATCC® CRL-3216™) were purchased from ATCC (Manassas, VA, USA). Cells were incubated in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% penicillin-streptomycin solution at 37 °C with 5% CO 2 .

Li W., Wu S., Xu H., Zhao X., Pan Y., Huang H., Lv H., Zhu X, & Liu Y. (2022). Novel variant in BRAT1 with the lethal neonatal rigidity and multifocal seizure syndrome. Pediatric research, 91(3).

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Cellular Uptake of Upconversion Nanoparticles

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HEK293T (ATCC^® CRL-3216^™, obtained from American Type Culture Collection, VA, US) cells were grown to confluence in eight-well chamber slides and incubated with 1 mg/mL of UCNPs and incubated at 37 °C for 10–60 min. At the end of incubation, the supernatant was replaced with fresh media and the cells were imaged using a confocal microscope to observe the localization of UCNPs. 2D images and 3D Z-stack images were obtained using the same.

Mei Q., Bansal A., Jayakumar M.K., Zhang Z., Zhang J., Huang H., Yu D., Ramachandra C.J., Hausenloy D.J., Soong T.W, & Zhang Y. (2019). Manipulating energy migration within single lanthanide activator for switchable upconversion emissions towards bidirectional photoactivation. Nature Communications, 10, 4416.

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HIV-1 Virus Infection Assay in TZM-bl Cells

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TZM-bl cells ^{100 (link)}, HIV-1 IIIB virus ^{101 –103 (link)}, HIV-1 IIIB (A17 variant) virus ^{104 (link)}, and the NNRTI resistant mutants (p5485, pNLGRINFQ, and p7324–1, all based on strain NL4–3) ^{105 (link)} were obtained from the NIH HIV Reagent Program. H9 [derivative of HuT 78] cells (ATCC® HTB176™) and 293T cells (ATCC® CRL-3216™) were obtained from the American Type Culture Collection. The IIIB and A17 viruses were grown in H9 cells to high titer, which was quantified using a HIV p24 (high sensitivity) AlphaLISA Detection Kit (PerkinElmer, Waltham, MA). The p5485, pNLGRINFQ, and p7324–1 virus plasmids were transfected into 293T cells using FuGene6 (Promega, Madison, WI) and then grown to high titer in H9 cells. The viruses were concentrated using Lenti-X Concentrator as needed (Takara Bio USA, Inc., Mountain View, CA). Infections of TZM-bl cells with these viruses was facilitated with 15 µg/mL DEAE-dextran (Sigma-Aldrich).

Lane T., Makarov V., Nelson J.A., Meeker R.B., Sanna G., Riabova O., Kazakova E., Monakhova N., Tsedilin A., Urbina F., Jones T., Suchy A, & Ekins S. (2023). N-phenyl-1-(phenylsulfonyl)-1H-1,2,4-triazol-3-amine as a new class of HIV-1 non-nucleoside reverse transcriptase inhibitor. Journal of medicinal chemistry, 66(9), 6193-6217.

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Culturing Transformed Brain Endothelial Cells

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The simian virus 40 large T antigen-transformed human brain microvascular endothelial cells (HBMEC) were cultured as previously described^{56 (link)–58}. Briefly, HBMECs were cultured in RPMI-1640 medium (Gibco Life Technologies, Karlsruhe, Germany) supplemented with 10% fetal calf serum (FCS, Gibco Life Technologies), 10% Nu serum^® IV (Corning, NY, USA 10%; Becton Dickinson), 1% sodium pyruvate (1 mM), 1% L-glutamine (2 mM), 1% non-essential amino acids (all purchased from GE Healthcare, Little Chalfont, UK), 5 U ml⁻¹ heparin (Biochrom, Berline, Germany) and 30 µg mL⁻¹ endothelial cell growth supplement (ECGS, CellSystems, Troisdorf, Germany). Cultures were incubated in a humid atmosphere at 37 °C with 5% CO₂. Cells between the 10th and 25th passages were used for apoptosis analysis. HBMEC were cultured in T25 flasks (Corning Costar Corporation, Cambridge, MA, USA). The embryonic kidney cell line HEK293T and the hepatocellular carcinoma cell line HepG2 were purchased from the American Type Culture Collection, ATCC^® CRL-3216^™ and ATCC^® HB-8065^™, respectively. HEK293T and HepG2 were routinely cultured in Dulbecco’s modified Eagle’s medium (DMEM) + GlutaMAX^™-I (Gibco Life Technologies, Karlsruhe, Germany) supplemented with 10% FCS.

Becam J., Walter T., Burgert A., Schlegel J., Sauer M., Seibel J, & Schubert-Unkmeir A. (2017). Antibacterial activity of ceramide and ceramide analogs against pathogenic Neisseria. Scientific Reports, 7, 17627.

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Lentiviral-mediated p53 Knockdown

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p53 knockdown was performed using four lentiviral-based shRNA constructs (Sigma-Aldrich, St. Louis, MO, USA). The shRNA p53-target sequences were as follows: p53si-2 (D3), 5′-CAC CATCCACTACAACTACAT-3′; p53si-3 (C12), 5′-CGGCGC ACAGAGGAAGAGAAT-3′; p53si-4 (E1), 5′-GAGGGATGT TTGGGAGATGTA-3′; p53si-5 (E2), 5′-GTCCAGATGAAG CTCCCAGAA-3′ and non-specific (NS), 5′-CAACAAGAT GAAGAGCACCAA-3′. Lentiviral stocks were generated by co-transfecting the HEK-293T cells (ATCC^® CRL-3216™; American Type Culture Collection, Manassas, VA, USA) with the plasmid vector, the psPAX2 packaging plasmids (Addgene plasmid 12260) and pMD2G envelope plasmid (Addgene plasmid 12259) (Addgene, Inc., Cambridge, MA, USA) using Lipofectamine 2000 (Invitrogen, Life Technologies, Carlsbad, CA, USA) according to the manufacturer's recommendations. The knockdown was verified by western blot analysis.

VOON Y.L., AHMAD M., WONG P.F., HUSAINI R., NG W.T., LEONG C.O., LANE D.P, & KHOO A.S. (2015). Nutlin-3 sensitizes nasopharyngeal carcinoma cells to cisplatin-induced cytotoxicity. Oncology Reports, 34(4), 1692-1700.

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