Skgt4

Manufactured by Merck Group

Sourced in United States

The SKGT4 is a laboratory equipment product from Merck Group. It is designed for general laboratory use. The core function of the SKGT4 is to assist in various experimental and analytical procedures conducted in a research or testing environment.

Automatically generated - may contain errors

Lab products found in correlation

Flo 1, by Merck Group (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) 293ft cell, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) 5 fluorouracil, by Merck Group (1 mentions) Sk gt 4, by Thermo Fisher Scientific (1 mentions) Rpmi medium, by Thermo Fisher Scientific (1 mentions) Caspase 3, by Cell Signaling Technology (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Dmem medium, by Thermo Fisher Scientific (1 mentions) Iq sybr green supermix, by Bio-Rad (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Het 1a cell, by American Type Culture Collection (1 mentions) Keratinocyte medium 2, by Cambrex (1 mentions) Begm bullet kit, by Cambrex (1 mentions) Epicm 2 medium, by ScienCell (1 mentions) Heepic, by ScienCell (1 mentions)

8 protocols using skgt4

Esophageal Cancer Cell Line Cultivation

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BAY1143572 was purchased from Active Biochemical (Wan Chai, Hong Kong).
5-fluorouracil and the human EAC cell lines OE33, FLO-1, and SKGT4 were
purchased from Sigma-Aldrich (St. Louis, MO). OE19, SKGT2, and ESO-26 cells were
obtained from Dr. Steven H. Lin (MD Anderson Cancer Center). OE33, OE19, SKGT2,
and ESO-26 cells were maintained in RPMI medium containing 2 mM L-glutamine and
10% fetal bovine serum (FBS). FLO-1 and SKGT4 cells were maintained in
Dulbecco’s modified Eagle’s medium (DMEM) containing 10% FBS. 293FT cells were
obtained from Invitrogen (Carlsbad, CA) and maintained in DMEM supplemented with
10% FBS and 500 µg/ml G418. All cell lines were maintained in a 5%
CO₂ atmosphere at 37°C and passaged at 80% confluence using 0.05%
trypsin−ethylenediaminetetraacetic acid for 3−5 min.

Tong Z., Mejia A., Veeranki O., Verma A., Correa A.M., Dokey R., Patel V., Solis L.M., Mino B., Kathkuda R., Rodriguez-Canales J., Lin S.H., Krishnan S., Kopetz S., Blum M., Ajani J.A., Hofstetter W.L, & Maru D.M. (2019). Targeting CDK9 and MCL-1 by a new CDK9/p-TEFb inhibitor with and without 5-fluorouracil in esophageal adenocarcinoma. Therapeutic Advances in Medical Oncology, 11, 1758835919864850.

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Esophageal Cancer Cell Lines Protocol

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BAY1143572 was purchased from Active Biochemical (Wan Chai, Hong Kong). Human esophageal adenocarcinoma cell lines, FLO1, OE33, and SKGT4 were purchased from Sigma Aldrich or ATCC. Radiation resistant esophageal adenocarcinoma cells (OE33R) were provided by our institutional Center for Radiation Oncology Research. Complete cell line information is provided in the Supplementary Materials and Methods.

Veeranki O.L., Tong Z., Dokey R., Mejia A., Zhang J., Qiao Y., Singh P.K., Katkhuda R., Mino B., Tailor R., Canales J.R., Bassett R., Ajani J., Wu J.Y., Kopetz S., Blum M., Hofstetter W., Tetzlaff M., Krishnan S., Lin S.H, & Maru D. (2019). Targeting cyclin-dependent kinase 9 by a novel inhibitor enhances radiosensitization and identifies Axl as a novel downstream target in esophageal adenocarcinoma. Oncotarget, 10(45), 4703-4718.

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Esophageal Adenocarcinoma Cell Lines: Cultivation and Characterization

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The human esophageal adenocarcinoma cell lines (OE33 and FLO-1) were kindly provided by Dr. David Beer (University of Michigan, Ann Arbor, MI). OE19 and SK-GT4 cells were obtained from Sigma-Aldrich (St. Louis, MO). OE33 and OE19 cells were maintained in RPMI medium (GIBCO, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS) (Invitrogen Life Technologies, Carlsbad, CA) and 1% penicillin/streptomycin (GIBCO). SK-GT-4 and FLO-1 cells were maintained in DMEM medium (GIBCO) supplemented with 10% FBS and 1% penicillin/streptomycin. The immortalized BART (non-dysplastic BE) was a kind gift from Dr. Rhonda Souza at University of Texas Southwestern whereas CPA (non-dysplastic BE) and CPB (BE with high-grade dysplasia) cells were purchased from ATCC (Manassas, VA); all cells were grown in accordance with supplier conditions. APE1, p-H2AX (S139), H2AX, caspase-3, PARP, and β-actin antibodies were obtained from Cell Signaling Technology (Danvers, MA). p-JNK (T183/Y185), JNK, p-p38 (T180/Y182), and p38 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). 8-OH-dG monoclonal antibody (clone 2E2) was purchased from Trevigen Inc., Gaithersburg, MD.

Hong J., Chen Z., Peng D., Zaika A., Revetta F., Washington M.K., Belkhiri A, & El-Rifai W. (2016). APE1-mediated DNA damage repair provides survival advantage for esophageal adenocarcinoma cells in response to acidic bile salts. Oncotarget, 7(13), 16688-16702.

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Esophageal Adenocarcinoma Cell Lines

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Esophageal adenocarcinoma cells OE33, FLO-1 and SKGT4 were purchased from Sigma-Aldrich (St. Louis, MO). ESO51 and KYAE-1 cells were obtained from Culture Collections (Public Health England, UK). OE33, ESO51 cells were maintained in a RPMI medium containing 2 mM of L-Glutamine, 10% of fetal bovine serum (FBS), 100 units/ml penicillin and 100 μg/ml of streptomycin. KYAE-1 cells were maintained in a medium of RPMI + Hams F12 (1:1) and FLO-1 and SKGT4 cells were maintained in a DMEM medium containing 10% of FBS, 100 units/ml penicillin and 100 μg/ml of streptomycin. 293 FT cells were obtained from Invitrogen (Carlsbad, CA) and maintained in a DMEM medium supplemented with 10 % FBS and 500μg/ml G418. Normal esophageal epithelial cells, HET-1A were provided by Dr. Xu (MD Anderson Cancer Center, Houston, TX) and maintained in a KSF medium (Lonza Walkersville Inc., Walkersville, MD). Cells were maintained in a 5% CO₂ atmosphere at 37°C and passaged at 80% confluence using 1 mM EDTA-0.025% trypsin for 3 to 5 minutes. All cell lines were authenticated by cell line validation core facility of UT M.D. Anderson Cancer Center.

Tong Z., Chatterjee D., Deng D., Veeranki O., Mejia A., Ajani J.A., Hofstetter W., Lin S., Guha S., Kopetz S., Krishnan S, & Maru D. (2017). Antitumor effects of cyclin dependent kinase 9 inhibition in esophageal adenocarcinoma. Oncotarget, 8(17), 28696-28710.

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Culturing Esophageal Carcinoma Cell Lines

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Esophageal carcinoma cell lines OE19 and SK-GT-4 (Sigma, USA) were cultured in 75 ml flasks at 37 °C with 5% CO₂, using Dulbecco’s Modified Eagle Medium supplemented with penicillin at 100/ml, streptomycin at 100 μg/ml and10% heat-inactivated fetal bovine serum (FBS) (Invitrogen, USA). The cell lines used in our study are validated with short tandem repeat (STR) profile analysis.

Zhu G., Zhang L., Dan J, & Zhu Q. (2020). Differential effects and mechanisms of local anesthetics on esophageal carcinoma cell migration, growth, survival and chemosensitivity. BMC Anesthesiology, 20, 126.

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Itraconazole Regulates BMP4 Expression

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The esophageal adenocarcinoma human cell line, SKGT4 (Sigma Chemical, St Louis, MO), was grown in culture medium containing 10% fetal bovine serum (Invitrogen, San Diego, CA). All cultures were maintained at 37°C in an incubator supplemented with 5% CO₂. Itraconazole (Sinoway Industrial Co, Xiamen, China) was dissolved in dimethyl sulfoxide (DMSO) for invitro experiments and was delivered to cells to reach concentrations of 1, 5, and 10μM in 2mL of cell culture medium. Total RNA was extracted from SKGT4 cells using TRIzol (Invitrogen, Frederick, MD) according to the manufacturer’s protocol. Total RNA of 500 ng was used for reverse transcription with high-capacity cDNA Reverse Transcription Kits (Applied Biosystems, Waltham, MA). Real-time PCR was performed using iQ SYBR Green Supermix (BioRad, Hercules, CA). The primer sequences were as follows: BMP4, 5′-GGC TGG AAT GAC TGG ATT GT-3′ and 5′-TGG TTG AGT TGA GGT GGT CA-3′; GAPDH, 5′-CAG CCT CAA GAT CAT CAG CA- 3′and 5′-TGT CGT CAT GAG TCC TTC CA-3′. Fold change in expression of target mRNA (BMP4) relative to GAPDH mRNA was calculated based on the threshold cycle (C_T) for amplification as 2^Δ(ΔCT), where ΔC_T =C_T,target – C_T,GAPDH.

Kelly R.J., Ansari A.M., Miyashita T., Zahurak M., Lay F., Ahmed A.K., Born L.J., Pezhouh M.K., Salimian K.J., Ng C., Matsangos A.E., Stricker-Krongrad A.H., Mukaisho K.I., Marti G.P., Chung C.H., Canto M.I., Rudek M.A., Meltzer S.J, & Harmon J.W. (2021). Targeting the Hedgehog Pathway Using Itraconazole to Prevent Progression of Barrett’s Esophagus to Invasive Esophageal Adenocarcinoma. Annals of surgery, 273(6), e206-e213.

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Culturing Esophageal Cell Lines

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Cell culture was similar to those we described previously[29 (link),32 (link),33 (link)]. Briefly, human esophageal squamous HET-1A cells were purchased from ATCC, Manassas, VA in 2011 and cultured in the bronchial epithelial cell medium (BEGM BulletKit, Cambrex, East Rutherford, NJ). Human Barrett’s cell line CP-A and Barrett’s dysplastic cell line CP-D were bought from ATCC (Manassas, VA) and cultured in Barrett’s medium containing keratinocyte medium-2 (Cambrex, Rockland, ME), 1.8 mmol/L CaCl₂, 5% fetal bovine serum, 400 ng/mL hydrocortisone, 20 ng/ml epidermal growth factor, 0.1 nmol/L cholera toxin, 20 μg/mL adenine, 5 μg/mL insulin, 70 μg/mL bovine pituitary extract, and antibiotics. EA cell line SK-GT-4 was purchased from Sigma and cultured in the Barrett’s medium.

Marketkar S., Li D., Yang D, & Cao W. (2017). TGR5 expression in benign, preneoplastic and neoplastic lesions of Barrett’s esophagus: Case series and findings. World Journal of Gastroenterology, 23(8), 1338-1344.

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Culturing Esophageal Cancer Cell Lines

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Human EC cell lines (SKGT4, OE33, and FLO-1) were obtained from Sigma-Aldrich. Normal esophageal epithelial cells (HEEPiC) was obtained from ScienCell Research Laboratories. The cell lines were identified by PCRamplified short tandem repeat analysis. Human EC cell line JHEso-Ad1 was a gift from Johns Hopkins University School of Medicine. EC cell lines were cultured in DMEM medium (Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin-streptomycin (Gibco). HEEPiC was cultured in EpiCM-2 medium (#4121, ScienCell). All cells were incubated at 37 °C with 5% CO 2 .

Wang Z., Ma K., Cheng Y., Abraham J.M., Liu X., Ke X., Wang Z, & Meltzer S.J. (2019). Synthetic circular multi-miR sponge simultaneously inhibits miR-21 and miR-93 in esophageal carcinoma. Laboratory investigation; a journal of technical methods and pathology, 99(10).

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