Mccoy s 5a

Manufactured by Lonza

Sourced in United States, Switzerland

The McCoy's 5A is a laboratory equipment product designed for general laboratory applications. It provides a stable and consistent work environment for various experimental procedures. The core function of the McCoy's 5A is to maintain a controlled temperature and humidity level within the workspace.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) Rpmi 1640, by Lonza (3 mentions) Rpmi 1640, by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Lonza (3 mentions) Dulbecco modified eagle medium (dmem), by Lonza (2 mentions) L glutamine, by Merck Group (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Lncap, by American Type Culture Collection (1 mentions) Cho cells, by American Type Culture Collection (1 mentions) Stemmacs msc expansion medium, by Miltenyi Biotec (1 mentions) Ht 29, by American Type Culture Collection (1 mentions) Roswell park memorial institute (rpmi), by Lonza (1 mentions) Hanks balanced salt solution (hbss), by Lonza (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) Blasticidin, by Merck Group (1 mentions) Lenticas9 blast, by Addgene (1 mentions) Hek293, by American Type Culture Collection (1 mentions) Mem vitamin solution, by Thermo Fisher Scientific (1 mentions) Pencillin streptomycin, by Merck Group (1 mentions)

Spelling variants of this product

McCoy’s 5A medium (25 citations) McCoy’s 5A Medium Modified (10 citations) McCoy 5A (4 citations) McCoy’s 5A Media (2 citations)

13 protocols using mccoy s 5a

Cytotoxicity Assays Using Cell Lines

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All chemicals, unless otherwise stated, were purchased from Sigma (St. Louis, MO, USA). Recombinant TRAIL (rTRAIL) was from R&D Systems (Minneapolis, MN, USA). Docetaxel, AKTi (MK-2206) and PI3Ki (LY294002) were from Stratech (Stratech Scientific, Ely, UK).
The colorectal carcinoma cell lines Colo205 (obtained from Dr. Eva Szegezdi) and HT-29 (ATCC, Manassas, VA, USA) were cultured in RPMI (Lonza, Basel, Switzerland) and in McCoy’s 5A (Lonza), respectively. HEK293 cells were grown in DMEM (Lonza) and CHO cells (ATCC) in Ham’s F12. The prostate cancer cell lines, LNCaP (ATCC), C4-2B (obtained from Dr. Wafa Al-Jamal) and PC3 (ATCC), were cultured in RPMI medium (Lonza). All media were supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, Waltham, MA, USA), 100 U/mL penicillin and 100 μg/mL streptomycin.
Human bone marrow derived MSCs were from Lonza, human adipose derived stem cells were from Amsbio (Cambridge, MA, USA) and human umbilical cord derived MSCs were from Promocell (Heidelberg, Germany). The different human MSCs were cultured in StemMACS MSC Expansion Medium (Miltenyi Biotec, Bergisch Gladbach, Germany). Murine MSCs were isolated and cultured as previously described [75 (link)]. In experiments where it was stated that MSCs were used, but not further defined, human bone marrow derived MSCs were used.

Mohr A., Chu T., Brooke G.N, & Zwacka R.M. (2019). MSC.sTRAIL Has Better Efficacy than MSC.FL-TRAIL and in Combination with AKTi Blocks Pro-Metastatic Cytokine Production in Prostate Cancer Cells. Cancers, 11(4), 568.

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Culturing Ovarian and Osteosarcoma Cells

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Ovarian carcinoma (2008) sensitive and (C13) cisplatin-resistant cells were grown in Roswell Park Memorial Institute medium (RPMI 1640) (Lonza) and osteosarcoma U2OS and U2OS-PT cells were grown in McCoy’s 5 A (Lonza) medium, both media were supplemented with 10% fetal bovine serum (FBS), 4 mM glutamine, 100 U/ml penicillin and 100 µg/ml streptomycin, in humidified condition at 5% CO₂ and 37 °C. For starvation treatment, cells were washed four times and then incubated in Hanks balanced salt solution (HBSS, Lonza) supplemented with 10 mM HEPES at pH 7.4, at 37 °C for the indicated time.

Vianello C., Cocetta V., Catanzaro D., Dorn GW I.I., De Milito A., Rizzolio F., Canzonieri V., Cecchin E., Roncato R., Toffoli G., Quagliariello V., Di Mauro A., Losito S., Maurea N., Scaffa C., Sales G., Scorrano L., Giacomello M, & Montopoli M. (2022). Cisplatin resistance can be curtailed by blunting Bnip3-mediated mitochondrial autophagy. Cell Death & Disease, 13(4), 398.

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Cell Culture Conditions for Various Cell Lines

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HCT116 cells were cultured in McCoy’s 5A (Lonza), RT112 and COLO205 cells were cultured in RPMI1640 (Sigma), PANC-1 and HEK 293 (all from ATCC) were cultured in DMEM (Sigma), and SKOV3 cells (NCI) were cultured in RPMI1640 supplemented with 1 mM sodium pyruvate (Gibco), 1% MEM NEAA (Gibco), and 1% MEM Vitamin solution (Gibco). Media were supplemented with 10% FBS (Sigma), 1% pencillin/streptomycin (Sigma) and 1% l-glutamine (Sigma). Cells were cultured at 37 °C and 5% CO₂ in a humidified incubator. Cells were STR authenticated and tested for mycoplasma regularly (Invivogen). HCT116-Cas9 cell line was generated by lentiviral transduction with lentiCas9-Blast (Addgene) and selection with 4 μg/mL of blasticidin (Sigma) for 3 days. KPC (Pdx1-cre;Kras^LSL.G12D/+;Tp53^LSL.R172H/+) and KPCZ (Pdx1-cre;Kras^LSL.G12D/+;Tp53^LSL.R172H/+; ZEB^fl/fl) are mouse pancreatic cancer cells without and with ZEB1 knockout [26 (link)] and were cultured in DMEM. TNF-alpha was from Gibco, and Cdk inhibitor (CGP-60474) from Tocris.

Gollavilli P.N., Parma B., Siddiqui A., Yang H., Ramesh V., Napoli F., Schwab A., Natesan R., Mielenz D., Asangani I.A., Brabletz T., Pilarsky C, & Ceppi P. (2021). The role of miR-200b/c in balancing EMT and proliferation revealed by an activity reporter. Oncogene, 40(12), 2309-2322.

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Cell culture protocols for various cell lines

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HCT116 cells were cultured in McCoy's 5A (Lonza) with 10% foetal bovine serum (FBS Sigma), 1% penicillin and streptomycin. HCT116 T300A cells were kindly provided by Dr. David Boone (Indiana University School of Medicine). J774.A1, HEK293, MDCK (a kind gift from Dr. P. Digard) and mouse embryonic fibroblast (MEF) cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco Life Technologies) containing 10% foetal bovine serum (FBS Sigma), 1% penicillin and streptomycin. Atg16L1^−/− MEFs were provided by Dr. Shizuo Akira (Osaka University). MCF10A cells were cultured in DMEM/F12 (Gibco) containing 5% horse serum (Sigma), EGF (20 ng/ml), hydrocortisone (0.5 mg/ml), cholera toxin (100 ng/ml) and insulin (10 μg/ml). All cells were maintained in an incubator at 37°C with 5% CO₂.

Fletcher K., Ulferts R., Jacquin E., Veith T., Gammoh N., Arasteh J.M., Mayer U., Carding S.R., Wileman T., Beale R, & Florey O. (2018). The WD40 domain of ATG16L1 is required for its non‐canonical role in lipidation of LC3 at single membranes. The EMBO Journal, 37(4), e97840.

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Semisynthetic Oleanolic Acid Derivatives

Cited 1 time

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Oleanolic acid was isolated from an industrial by-product obtained during the process of mistletoe herb essence production. Spectral data of the resulting chemicals were consistent with the data from the literature [63 ]. The semisynthetic OA derivatives, methyl 3-hydroxyimino-11-oxoolean-12-en-28-oate (HIMOXOL) and 12α-bromo-3-hydroxyimonoolean-28→13-olide (Br-HIMOLID) were synthesized as described in the earlier studies [64 (link),65 (link)] at the Department of Organic Chemistry, Poznan University of Medical Sciences, Poland (Figure 8). Before use in the experiment, the derivatives were dissolved in DMSO and stored at 4 °C.
The cell culture medium, McCoy’s 5A, and fetal bovine serum were purchased from Lonza (Lonza, Verviers, Belgium). Rapamycin, DMSO, camptothecin, propidium iodide, RNAse, and MTT were obtained from Sigma-Aldrich (Sigma-Aldrich, Munich, Germany). Gefitinib was purchased from Cell Signaling Technology (Cell Signaling Technology, Danvers, MA, USA).

Lisiak N.M., Lewicka I., Kaczmarek M., Kujawski J., Bednarczyk-Cwynar B., Zaprutko L, & Rubis B. (2021). Oleanolic Acid’s Semisynthetic Derivatives HIMOXOL and Br-HIMOLID Show Proautophagic Potential and Inhibit Migration of HER2-Positive Breast Cancer Cells In Vitro. International Journal of Molecular Sciences, 22(20), 11273.

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Bladder Cancer Cell Line Characterization and Spiking

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HT-1197 and RT4 bladder cancer cell lines were purchased from ATCC (Manassas, USA) and their authenticity was verified using the STR DNA profiling with the StemElite™ ID System kit (Promega, Madison, WI, USA) by INT Genomics Core Facility. HT-1197 and RT4 cells were cultured in RPMI 1640 and McCoy's 5A media (Lonza, Slough, UK), respectively, supplemented with 10% South America origin Fetal Bovine Serum (Lonza), in humidified 5% CO₂ atmosphere.
FACS analysis for the expression of cell surface EPCAM (mouse IgG1κ anti- human EPCAM-PerCP-Cy7 antibody, clone 1B7, EBiosciences, San Diego, USA) and ErbB2 (mouse IgG2B anti- human ErbB2 antibody, clone 191924, R&D Systems, Minneapolis, MN, USA) was performed on fresh monodisperse RT4 cell suspensions versus isotype control samples.
For spiking experiments, highly diluted cell suspensions were prepared in culture dishes and single viable cells (detectable by Trypan blue exclusion assay) were micropipetted under an inverted optical microscope directly into conical tubes containing 5 mL of whole blood from healthy volunteer. Spiked-in samples were stored at 4°C in the dark for no more than 1 hour before processing.

Fina E., Necchi A., Bottelli S., Reduzzi C., Pizzamiglio S., Iacona C., Daidone M.G., Verderio P, & Cappelletti V. (2017). Detection of Circulating Tumour Cells in Urothelial Cancers and Clinical Correlations: Comparison of Two Methods. Disease Markers, 2017, 3414910.

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Apoptosis Analysis of Cisplatin-Treated Cell Lines

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HeLa cells, purchased from DSMZ GmbH (Leibniz, Germany), and an ME-180 (HTB-33) cell line, purchased from ATCC (Manassas, VA, USA), were cultured in RPMI 1640 (Gibco, ThermoFischer, Waltham, MA, USA) and McCoy’s 5A (Lonza, Switzerland) supplemented with 2 mM L-glutamine, 10% fetal bovine serum (FBS) (Gibco, ThermoFischer, Waltham, MA, USA) in a humidified atmosphere of 5% CO₂ at 37 °C. Cisplatin (Santa Cruz Biotechnology, Dallas, TX, USA) treatments were carried out in triplicates essentially as described previously [12 (link)]. 0.1% (v/v) DMSO was used as a negative control for Cisplatin. Treated cells were trypsinized by 1X Trypsin-EDTA (Gibco, ThermoFischer, Waltham, MA, USA) and washed in 1X cold PBS (Gibco, ThermoFischer, Waltham, MA, USA). Cell pellets were stained with Annexin V-PE (BD Biosciences, Franklin Lakes, NJ, USA) and 7AAD (BD Biosciences, Franklin Lakes, NJ, USA) in the presence of 1X Annexin binding buffer (BD Biosciences, Franklin Lakes, NJ, USA) followed by incubation at room temperature for 15 min in the dark. The rates of Annexin V and/or 7AAD-positive cells were quantified by a FACSCanto flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA).

Alasar A.A., Tüncel Ö., Gelmez A.B., Sağlam B., Vatansever İ.E, & Akgül B. (2022). Genomewide m6A Mapping Uncovers Dynamic Changes in the m6A Epitranscriptome of Cisplatin-Treated Apoptotic HeLa Cells. Cells, 11(23), 3905.

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Cell Viability and Apoptosis Assays

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Alpha-MEM, McCoy’s 5A and RPMI-1640 mediums, fetal bovin serum (FBS), penicillin and streptomycin were from Lonza (Lonza Milano SRL, Milan, Italy). Torin-2, RAD001, and MK-2206 were from Selleck Chemicals (Houston, TX, USA). For cell viability determination, Cell Proliferation Kit I (MTT) was purchased from Roche Applied Science (Basel, Switzerland). Annexin V/7-AAD detection kit was from Merck-Millipore (Darmstadt, Germany). Akt-1, Ser 473 p-Akt-1, and FoxO3A primary antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA) while all the other antibodies were from Cell Signaling Technology (Danvers, MA, USA), including the rabbit secondary antibody. The mouse secondary antibody, Bafilomycin A1, Z-VAD-fmk, 3-Methyladenine (3-MA), 1,4-Diazabicyclo[2.2.2]octane (DABCO) and 4′, 6 diamidino-2-pheny-lindole (DAPI) were from Sigma Aldrich (Milan, Italy). Signals were detected with the ECL Plus reagent purchased from Perkin Elmer (Boston, MA, USA).

Simioni C., Cani A., Martelli A.M., Zauli G., Tabellini G., McCubrey J., Capitani S, & Neri L.M. (2014). Activity of the novel mTOR inhibitor Torin-2 in B-precursor acute lymphoblastic leukemia and its therapeutic potential to prevent Akt reactivation. Oncotarget, 5(20), 10034-10047.

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Cell Culture Maintenance Protocols

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Caki-1 cells were maintained in McCoy’s 5A (Lonza, Basel, Switzerland) medium, 786-O cells were maintained in RPMI 1640 (Lonza) medium, A-498 cells were cultured in EMEM (Minimum Essential Medium Eagle, Lonza) medium, and HEK293 cells were cultured in DMEM (Dulbecco’s Modified Eagle’s Medium, Lonza) medium. All media were supplemented with 10% FBS (Fetal Bovine Serum, Sigma-Aldrich, St. Louis, MO, USA), 1% L-Glutamine (Sigma-Aldrich), and 1% penicillin/streptomycin (Sigma-Aldrich). Cells were incubated at 37 °C and 5% CO₂.

Emaldi M, & Nunes-Xavier C.E. (2022). B7-H4 Immune Checkpoint Protein Affects Viability and Targeted Therapy of Renal Cancer Cells. Cells, 11(9), 1448.

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Isolation and Culture of GCB Cells

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Cell lines used throughout the manuscript are listed in Supplementary Table S1, including their origin and culture conditions. Cell lines were either cultured in RPMI 1640 (Gibco, Waltham, MA, USA; Lonza BioWhittaker, Walkersville, MD, USA in case of ST486), McCoys5A (Lonza BioWhittaker, Walkersville, MD, USA) or DMEM (Lonza BioWhittaker, Walkersville, MD, USA), supplemented with 10–20% fetal bovine serum (Cambrex Biosciences, Walkersville, MD, USA). All media were supplemented with 100 units/mL of penicillin, 100 µg/mL of streptomycin and 2 mM of glutamine (Cambrex Biosciences, Walkersville, MD, USA). The origin of the cell lines was confirmed with STR DNA analyses on a regular basis and mycoplasma tests were performed to exclude contamination. The cultures were maintained at 37 °C in a humidified air atmosphere supplemented with 5% CO₂.
GCB-cells (defined as CD20+IgD−CD38+) were sorted from routinely removed tonsil specimens as described previously [13 (link)]. Written permission for the use of the tonsil tissues to isolate GCB-cells was obtained from the parents of the children. The study protocol was consistent with international ethical guidelines (the Declaration of Helsinki and the International Conference on Harmonization Guidelines for Good Clinical Practice).

Ziel-Swier L.J., Liu Y., Seitz A., de Jong D., Koerts J., Rutgers B., Veenstra R., Razak F.R., Dzikiewicz-Krawczyk A., van den Berg A, & Kluiver J. (2022). The Role of the MYC/miR-150/MYB/ZDHHC11 Network in Hodgkin Lymphoma and Diffuse Large B-Cell Lymphoma. Genes, 13(2), 227.

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