Hep 2

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Hep-2 is a laboratory equipment used for the detection and identification of antinuclear antibodies (ANAs) in human serum samples. It is a cell-based indirect immunofluorescence assay (IFA) system that utilizes Hep-2 cells as the substrate. The Hep-2 system provides a standardized method for the screening and semi-quantitative analysis of ANAs, which are associated with various autoimmune disorders.

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HEPES (4332 citations) 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) (79 citations) N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) (32 citations) 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer (21 citations) 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), (7 citations) Hep-2 cells (3 citations) 4-(2-68 hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), (3 citations) 4-[2-hydroxyethyl]-1-piperazineerhanesulfonic acid (HEPES; (3 citations) Hepes buffer (pH 7.2) (2 citations) 4-(2-hydroxyethyl)-1-piperazinee-thanesulfonic acid (HEPES)-buffered saline solution (2 citations)

13 protocols using hep 2

Human Oral Cancer Cell Interactions

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The human oral squamous cell carcinoma cell lines Hep-2 and human lymphatic endothelial cells (hLECs) were obtained from BeNa Culture Collection (Hangzhou, Zhejiang, China). hLEC were cultured with 85% RPMI-1640 (Thermo Fisher Scientific, Waltham, MA, USA) and 15% FBS (Atlas Biologicals), and Hep-2 were maintained in RPMI-1640 (Thermo Fisher Scientific, Waltham, MA, USA) and 10% FBS at 37 °C in a humidified atmosphere containing 5% CO₂.
The hLEC cells were plated in a six-well plate and treated with EBM-2 or Hep-2 conditioned medium for 48 h. Then the cells were collected and used to extract protein and RNA for correlation detection (western blot analysis and qRT-PCR).

Zhang C., Zhu M., Wang W., Chen D., Chen S, & Zheng H. (2019). TNF-α promotes tumor lymph angiogenesis in head and neck squamous cell carcinoma through regulation of ERK3. Translational Cancer Research, 8(6), 2439-2448.

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Cultured Human Cancer Cell Lines

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Human breast cancer cells, MDA-MB-231 and MCF-7, were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, USA); human glioma cell line U251, human lung carcinoma cell line A549, human microvascular endothelial cell line HMEC-1, human esophageal cancer cell line EC109, laryngeal carcinoma cell line Hep2, human hepatocarcinoma cell line HepG2, human non-small cell lung cancer cell line NCI-H157, human lung cancer cell line NCI-H460, and prostate cancer cell line PC3 were maintained in Roswell Park Memorial Institute RPMI_1640 medium (Thermo Fisher Scientific). All media were supplemented with 10% fetal calf serum (Thermo Fisher Scientific), 10 U/mL penicillin, and 100 μg/mL streptomycin (Thermo Fisher Scientific). U251, EC109, Hep2, NCI-H157, and PC3 cell lines were obtained from the tumor cell library of the Peking Union Medical College (PUMC); MDA-MB-231, MCF-7, A549, HMEC-1, HepG2, and NCI-H460 cell lines were purchased from American Type Culture Collection (ATCC), Manassas, VA, USA.

He X., Hao Y., Long W., Song N., Fan S, & Meng A. (2015). Exploration of peptide T7 and its derivative as integrin αvβ3-targeted imaging agents. OncoTargets and therapy, 8, 1483-1491.

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Cultivation of HEp-2 Cervical Carcinoma Cells

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The human cervical carcinoma cell line HEp-2 (ATCC^® CCL-23™), derived from epithelial cells, was purchased from ATCC (Manassas, Virginia, USA) and maintained in liquid nitrogen. An aliquot was transferred to a 25-cm² plastic tissue culture flask (Corning Costar, Corning, Nueva York, USA) containing 3 ml EMEM (ATCC^® 30–2003™; ATCC) supplemented (EMEMsup) with 1% nonessential amino acids, 1% penicillin–streptomycin (GIBCO, Thermo Scientific, Waltham, Massachusetts, USA), and 10% fetal bovine serum (Corning). The HEp-2 cells were incubated at 37°C in 5% CO₂ (Steri-Cycle CO₂ Incubator; Thermo Scientific) and used between passages 7 and 30.

Ortiz Y., García-Heredia A., Merino-Mascorro A., García S., Solís-Soto L, & Heredia N. (2021). Natural and synthetic antimicrobials reduce adherence of enteroaggregative and enterohemorrhagic Escherichia coli to epithelial cells. PLoS ONE, 16(5), e0251096.

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Evaluation of Esculetin and STAT3 Modulators in Laryngeal Cancer

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Human laryngeal cancer cell lines Hep-2, TU-212, and M4e, and human tubular epithelial cell line HK2 were purchased from the American Type Culture Collection (Manassas, VA USA). The Hep-2, TU-212, and HK2 cells were cultured in RPMI Medium 1640 (Thermo Fisher, Waltham, MA, USA), and M4e cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco; Thermo Fisher, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA). Cells were cultured in a 37°C incubator containing 5% CO₂. When growth was in logarithmic phase, cells were seeded onto 96-well plates for further study.
Esculetin was purchased from Sigma-Aldrich (99.99% purity) as 100 mM stock solution, and cisplatin was also purchased from Sigma-Aldrich. STAT3 inhibitor C188-9 was purchased from Selleck Chemicals (Houston, TX, USA) and the STAT3 activator colivelin was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). In an in vitro study, Esc, C188-9, and cisplatin were serially diluted with RPMI Medium 1640 triple and triple. Final working concentrations were 0.0457, 0.1369, 0.4120, 1.229, 3.700, 11.10, and 33.29 μM and the highest working concentration was 100 μM.

Zhang G., Xu Y, & Zhou H.F. (2019). Esculetin Inhibits Proliferation, Invasion, and Migration of Laryngeal Cancer In Vitro and In Vivo by Inhibiting Janus Kinas (JAK)-Signal Transducer and Activator of Transcription-3 (STAT3) Activation. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 25, 7853-7863.

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Culturing Human Laryngeal Carcinoma Hep-2 Cells

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Human laryngeal carcinoma cells Hep-2 were obtained from BIOBAIYE (Shanghai China). Hep-2 cells were routinely cultured in DMEM (Gibco), which consisted of 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin (Solarbio) at 37 °C. Placed within an incubator of 5% CO2, the cells were digested with 0.25% membrane protease (Sigma) every 2–3 days.

Wang X., Pan Y., Ou Y., Duan T., Zou Y, & Zhou X. (2022). Construction and validation of immune-related LncRNAs classifier to predict prognosis and immunotherapy response in laryngeal squamous cell carcinoma. World Journal of Surgical Oncology, 20, 164.

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Establishing Cisplatin-Resistant Cell Lines

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Nasopharyngeal epithelial cell line NP69 and laryngeal cancer lines including Hep-2 and TU-177 cell lines were purchased from ATCC. Hep-2 cells were cultured in Eagle's Minimum Essential Medium (EMEM) supplement with 10% fetal bovine serum (FBS, Gibco) at 37°C with 5% CO₂. TU-177 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplement with 10% FBS at 37°C with 5% CO₂.
Cisplatin-resistant Hep-2 cells (Hep-2/R) and TU-177 cells (TU-177/R) were established by the incubation of Hep-2 and TU-177 with stepwise increased concentrations of cisplatin as previously described method (22 (link)).

Zhang Y., Zhang H., Dong J., Zhao P., Hao F., Han H, & Bian Y. (2022). CAPRIN1 Enhances Chemoresistance and Glycolysis in Laryngeal Squamous Cell Carcinoma via Regulation of ZIC5. Journal of Oncology, 2022, 6160539.

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Culturing Human Laryngeal Cancer Cells

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The human laryngeal cancer cell line Hep2 was obtained from KeyGen Biotech Company (Nanjing, China). Hep2 cells were cultured in RPMI1640 (GIBCO Thermo Fisher Scientific, MA, USA) supplemented with 10% fetal bovine serum (FBS, Israel), 100 U/ml penicillin, and 100 mg/ml streptomycin (Solarbio, China) in an incubator at 37°C with 5% CO₂.

Wang P.P., Ding S.Y., Sun Y.Y., Li Y.H, & Fu W.N. (2021). MYCT1 Inhibits the Adhesion and Migration of Laryngeal Cancer Cells Potentially Through Repressing Collagen VI. Frontiers in Oncology, 10, 564733.

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Laryngeal Cancer Cell Lines Profiling

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Human laryngeal cancer cell lines, TU212 and M4e, human normal larynx epithelial HBE cells, and human normal liver HHL-5 cells, were purchased from the American Type Culture Collection (ATCC; Manassas, VA USA). The mouse normal larynx epithelial RTE cells, and human laryngeal cancer HEP-2 cells were purchased from the Nanjing KeyGen Biotech, Co., Ltd. (Nanjing, China). TU212, HEP-2 and RTE cells were routinely cultured in RPMI-1640 medium (Gibco, Carlsbad, CA, USA), containing 10% fetal bovine serum (FBS; Gibco), 1% penicillin/streptomycin. The cell lines M4e, HBE and HHL-5 were cultured in Dulbeccos modified Eagles medium (DMEM; Gibco) supplemented with 10% FBS, 100 U/ml penicillin and 100 µg/ml streptomycin. All cells were kept in a humidified atmosphere with 5% CO₂ and 95% humidity at 37°C in an incubator. Galangin (>98% purity), purchased from the Hangzhou DayangChem, Co., Ltd., (Hangzhou, China) was used for the treatment of human laryngeal cancer dissolved in dimethyl sulfoxide (DMSO) and then stored at −20°C for experimental treatment use. The final DMSO concentration in cells is <0.1% (v/v) in each treatment.

Wang H.X, & Tang C. (2017). Galangin suppresses human laryngeal carcinoma via modulation of caspase-3 and AKT signaling pathways. Oncology Reports, 38(2), 703-714.

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Culturing LSCC Hep-2 Human Epithelial Cells

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The LSCC cell line human epithelial-2 (Hep-2) was purchased from and authenticated by the Cell Bank of the Chinese Academy of Sciences (Shanghai, People's Republic of China), and maintained in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (Gibco, Grand Island, NY) in a humidified atmosphere containing 5% CO 2 at 37°C.

Zhang Z., Wang X., Cao S., Han X., Wang Z., Zhao X., Liu X., Li G., Pan X, & Lei D. (2018). The Long Noncoding RNA TUG1 Promotes Laryngeal Cancer Proliferation and Migration. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 49(6).

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Modulating circZNF609 and miR-134-5p in LSCC Cell Lines

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The human LSCC cell lines TU177, TU686, TU212, LSC-1 and Hep-2 were obtained from Shanghai Institute of Biological Science Cell Center. Hep-2 cells were cultured in F12K medium (Invitrogen), and the remaining cells were cultured in RPMI 1640 medium (Invitrogen).
Cell transfection was carried out when cells in culture reached 60–80% confluence using Lipofectamine® 2000. Small interfering RNAs (siRNAs) targeting circZNF609#1 (si-circZNF609#1, 5ʹ-GTCAAGTCTGAAAAGCAATGA-3ʹ), circZNF609#2 (5ʹ-TGCCCTAGTACTACCCTGCAT-3ʹ) and circZNF609#3 (5’-TTGACTGCATCGTAGCCAAAC-3’) and negative control (si-NC, 5’-UUCUCCGAACGUGUCACGUTT-3’) were purchased from Shanghai GenePharma Co., Ltd. The miR-134-5p mimetic, agomir and controls were purchased from Guangzhou RiboBio Co., Ltd. The transfection concentrations of oligonucleotides were as follows: si-NC, 40 nM; si- circZNF609, 40 nM; si-NC, 40 nM; si-EGFR, 40 nM; miR-134-5p mimetic, 50 nM; and miRNA control, agomir and miRNA control, 50 nM. Lipofectamine® 2000 Reagent and si-RNAs or miR-mimic were diluted with serum-free DMEM medium, mixed together and incubated at room temperature for 20 min. This solution was subsequently added to LSC-1 and Hep-2 cells for transfection at 37°C for 4–6 h in a humidified incubator containing 5% CO₂.

Yin X., Wang J., Shan C., Jia Q., Bian Y, & Zhang H. (2022). Circular RNA ZNF609 promotes laryngeal squamous cell carcinoma progression by upregulating epidermal growth factor receptor via sponging microRNA-134-5p. Bioengineered, 13(3), 6929-6941.

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