Uacc903

Manufactured by American Type Culture Collection

The UACC903 is a compact, high-performance cell culture incubator designed for laboratory use. It maintains a controlled environment for the growth and maintenance of cell cultures, providing precise temperature, humidity, and atmospheric gas regulation.

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Lab products found in correlation

Chl 1, by American Type Culture Collection (3 mentions) Sk mel 5, by American Type Culture Collection (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Sk mel 5, by Thermo Fisher Scientific (1 mentions) Mek1 2, by Merck Group (1 mentions) Sp600125, by Merck Group (1 mentions) Chl 1, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) U0126, by Merck Group (1 mentions) Sk mel 28, by American Type Culture Collection (1 mentions) Wm 239, by American Type Culture Collection (1 mentions) Wm164, by American Type Culture Collection (1 mentions) Gm12878, by Coriell Institute for Medical Research (1 mentions) Phenazine methosulfate pms, by Merck Group (1 mentions) T4 polynucleotide kinase, by Illumina (1 mentions) Mcf 7, by American Type Culture Collection (1 mentions) Panc 1, by American Type Culture Collection (1 mentions) γ 32p atp, by PerkinElmer (1 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions) Dntps, by Promega (1 mentions)

4 protocols using uacc903

Melanoma Cell Line Manipulation and Pharmacological Inhibition

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Five human melanoma cells lines, including A-375, 1205Lu, UACC903, CHL-1 and sk-mel-5, were purchased from American Type Culture Collection. Cells were maintained in RPMI-1640 medium (A-375, 1205Lu) or DMEM medium (UACC903, CHL-1 and sk-mel-5) respectively supplemented with 10% fetal bovine serum (Gibco, CA, USA) in a 37°C humidified atmosphere of 5% CO₂.
Knockdown of BANCR was induced by transfection with BANCR shRNA (Genepharma, Shanghai, China) using Lipofectamine2000 (Invitrogen, CA, USA). The target sequences were as follows: shRNA #1: 5′-CGGAAATAGACTGCAGCAC CAA-3′; shRNA #2: 5′-CCTTTATGGATTCAACTGTAAT-3′. Ectopic expression of BANCR was achieved through pCDNA3.1-BANCR transfection using Lipofectamine2000. Oligonucleotides for amplification of BANCR were as follows: sense: 5′- CAGGAAGGGGTGAATGAAGA-3′; antisense: 5′- CCAGTGCAGGGTAATGTGTG-3′.
Stable transfectants were selected in medium containing 600 µg/mL G418 (Invitrogen, CA, USA) and used in further assays or RNA/protein extraction.
For treatment with pharmacological inhibitors, the overnight starved cells were kept in complete medium containing 10 µM U0126 (inhibitor of the upstream ERK1/2 activator MEK1/2, Sigma-Aldrich, MO, USA) or 20 µM SP600125 (JNK inhibitor, Sigma-Aldrich, MO, USA). DMSO was used as a control. Medium containing inhibitor was renewed every day.

Li R., Zhang L., Jia L., Duan Y., Li Y., Bao L, & Sha N. (2014). Long Non-Coding RNA BANCR Promotes Proliferation in Malignant Melanoma by Regulating MAPK Pathway Activation. PLoS ONE, 9(6), e100893.

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Modulating melanoma cell lines via miR-143 and Syn-1

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Human melanoma cells lines, including A-375, 1205Lu, UACC903, CHL-1 and sk-mel-5, were purchased from American Type Culture Collection. Cells were maintained in RPMI-1640 medium (A-375, 1205Lu) or DMEM medium (UACC903, CHL-1 and sk-mel-5) respectively supplemented with 10% fetal bovine serum in a 37°C humidified atmosphere of 5% CO₂.
Ectopic expression of miR-143 in cells was obtained by transfection with miR-143 mimics or inhibitors (50 nM) using Lipofectamine2000. Knockdown of Syn-1 was performed using Syn-1 siRNA (100 nM). Human cDNA encoding Syn-1 was constructed using pcDNA 3.0 vector and transfected into cells to induce Syn-1's reconstitution. Cells were plated in 6-well or 96-well plates and transfected for 24 h or 48 h. Transfected cells were used in further assays or RNA/protein extraction.

Li R., Zhang L., Jia L., Duan Y., Li Y., Wang J., Bao L, & Sha N. (2014). MicroRNA-143 Targets Syndecan-1 to Repress Cell Growth in Melanoma. PLoS ONE, 9(4), e94855.

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Melanoma Cell Lines and Vemurafenib Resistance

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Human melanoma cell lines (A375, SK-mel-28, UACC903, MGH-MC-1, K4, WM239, WM1158, WM164, G-mel, CHL-1, SK-mel-119) were developed in-house, purchased from the American Type Culture Collection (Rockville, MD) or gifts from Meenhard Herlyn (Wistar Institute, Philadelphia, PA). Vemurafenib resistant melanoma cell lines (A375-P, SK-Mel 28-P, UACC903-P, MGH-MC-1-P, K4-P, WM239-P and WM1158-P) were obtained by sequentially exposing of cells to escalating doses of VEM in 2014. DMEM medium with 10% fetal bovine serum (FBS), supplemented with 100 units/ml penicillin, 100 μg/ml streptomycin, and 2 mM glutamine was used for routine culturing of cells; VEM-resistant cells were cultured in VEM-free media for 10–14 days prior to re-testing VEM sensitivity; this drug holiday period had no effect on the VEM resistance (data not shown). All cells were incubated in a humidified atmosphere of 95% air and 5% CO₂ at 37°C. Tumor specimens from patients were obtained prior to treatment with BRAFi (vemurafenib) or BRAFi+MEKi (dabrafenib+trametinib) and post-relapse as indicated in Table S1. Acquisition of tissue was covered under a protocol approved by the DF/Harvard Cancer Center (legacy #11-181; Boston, U.S.) in accordance with the Declaration of Helsinki.

Miao B., Ji Z., Tan L., Taylor M., Zhang J., Choi H.G., Frederick D.T., Kumar R., Wargo J.A., Flaherty K.T., Gray N.S, & Tsao H. (2014). EphA2 is a Mediator of Vemurafenib Resistance and a Novel Therapeutic Target in Melanoma. Cancer discovery, 5(3), 274-287.

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Cell Viability Assay Protocols

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T4 polynucleotide kinase was purchased from Epicenter (Lexington, KY) and [γ-³²P]ATP (6000 Ci/mmol) from PerkinElmer (Waltham, MA). DNA oligomers were purchased from IDT (Coralville, IA). The concentrations of the dNTPs (Promega, Madison WI) were each determined by UV absorbance.³⁵ HNK, BLM, and TMZ, purchased from Sigma-Aldrich (St. Louis, MO), were freshly prepared in DMSO, stored at −80 °C, and diluted in buffer or complete medium just before use. 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) was purchased from Promega (Madison, WI, USA), and phenazine metho-sulfate (PMS) was purchased from Sigma-Aldrich (St. Louis, MO, USA). MTS/PMS solution was prepared at a concentration of 2 mg/mL of MTS and 0.92 mg/mL of PMS in phosphate-buffered saline (PBS) and stored in dark bottle at 4 °C. Fetal bovine serum was purchased from Atlanta Biologicals Inc. The human cancer cell lines, MCF7, A549, PANC-1, UACC903, were purchased from American Type Culture Collection (ATCC), and GM12878 cells were purchased from Coriell Institute for Medical Research.

Prakasha Gowda A.S., Suo Z, & Spratt T.E. (2017). Honokiol Inhibits DNA Polymerases β and λ and Increases Bleomycin Sensitivity of Human Cancer Cells. Chemical research in toxicology, 30(2), 715-725.

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