Cryo nhmc

Manufactured by Cambrex

Sourced in United States

The Cryo NHMC is a laboratory instrument designed for the cryogenic cooling and temperature control of samples. It provides precise temperature regulation and maintains samples at low temperatures, enabling research and analysis in a wide range of scientific applications.

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Lab products found in correlation

6 protocols using cryo nhmc

Cell Culture and Reagent Procurement for Apoptosis Research

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Caki, ACHN, A498, MDA-MB231, and U251MG cells were purchased from the American Type Culture Collection (Manassas, VA, USA). The mouse kidney cells (TMCK-1) were a gift from Dr. T.J. Lee (Yeungnam University, Korea). A primary culture of human mesangial cells (Cryo NHMC) was purchased from Clonetics (San Diego, CA, USA). The cells were cultured in Dulbecco's modified Eagle medium that contained 10% fetal bovine serum, 20 mM Hepes buffer, and 100 μg/ml gentamicin. The PCR primers were obtained from Macrogen Inc. (Seoul, Korea), and the other chemicals were obtained from Sigma (St. Louis, MO, USA). NAC and Trolox were obtained from Calbiochem (San Diego, CA, USA). The anti-DR5, anti-Bcl-2, anti-Bcl-xL, anti-Mcl-1, anti-XIAP, and anti-PARP antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The anti-procaspase-3 and anticleaved caspase-3 antibody was obtained from Cell Signaling Technology (Beverly, MA, USA). The anti-FADD antibody was obtained from BD Bioscience (San Jose, CA, USA). The anti-c-FLIP(L) antibody was obtained from the ALEXIS Corporation (San Diego, CA, USA). The anti-Bim antibody was obtained from Millipore Corporation (Billerica, MA, USA). The anti-actin antibody was obtained from Sigma. The recombinant human TRAIL was obtained from KOMA Biotech (Seoul, Korea).

Min K.J., Seo B.R., Bae Y.C., Yoo Y.H, & Kwon T.K. (2014). Antipsychotic agent thioridazine sensitizes renal carcinoma Caki cells to TRAIL-induced apoptosis through reactive oxygen species-mediated inhibition of Akt signaling and downregulation of Mcl-1 and c-FLIP(L). Cell Death & Disease, 5(2), e1063-.

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TRAIL-induced Apoptosis in Kidney Cells

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Mouse kidney cells (TCMK-1) were a gift from Dr. T.J. Lee (Yeungnam University, Korea). Primary culture of human mesangial cells (Cryo NHMC) were purchased from Clonetics (San Diego, CA, USA), and other cell lines are from the ATCC (Manassas, VA, USA). All cells were maintained in Dulbecco’s modified Eagle’s medium, and supplemented with 10% FBS, 5% antibiotic solution, and 100 μg/mL gentamycin. Garcinol and lactacystin was purchased from Enzo Life Sciences (Ann Arbor, MI, USA). Recombinant human TRAIL and z-VAD-fmk were purchased from R&D (Minneapolis, MN, USA). Calbiochem supplied N-acetyl-l-cysteine (NAC), trolox, and MG132 (San Diego, CA, USA). The following antibodies were used: anti-XIAP, -Bcl-2, -Mcl-1, -survivin, -Cbl, and -Itch (Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-c-FLIP (ALEXIS Corporation, San Diego, CA, USA); anti-PARP, -DR5, -PSMA5, -PSMD4/S5a, -cleaved caspase-3 (Cell Signaling Technology, Beverly, MA, USA); anti-caspase-3 (Enzo Life Sciences, Ann Arbor, MI, USA); anti-DR4 (Abcam, Cambridge, MA, USA); anti-actin (Sigma-Aldrich, St. Louis, MO, USA). Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Bioneer (Daejeon, Korea) supplied DR5 siRNA and GFP siRNA (control siRNA), respectively. Sigma Chemical Co. supplied other reagents (St. Louis, MO, USA).

Kim S., Seo S.U., Min K.J., Woo S.M., Nam J.O., Kubatka P., Kim S., Park J.W, & Kwon T.K. (2018). Garcinol Enhances TRAIL-Induced Apoptotic Cell Death through Up-Regulation of DR5 and Down-Regulation of c-FLIP Expression. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(7), 1614.

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Assessing TRAIL-Induced Apoptosis in Cancer Cells

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Caki, MDA-MB-361 and U251MG cells were obtained from the American Type Culture Collection (ATCC) (Rockville, Maryland, USA). The culture medium used throughout these experiments was Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS), 20 mM HEPES buffer, and 100 mg/ml gentamycin. The mouse kidney cells, TMCK-1, were a gift from Dr T.J. Lee (Yeungnam University, Korea). Primary cultures of human mesangial cells (Cryo NHMC) and their corresponding growth medium (CC-3146 MsGM) were purchased from Clonetics (San Diego, California, USA). Recombinant human TRAIL and z-VAD-fmk were purchased from R&D system. CQ was purchased from Sigma Chemical Co. (St Louis, Missouri, USA). Anti-Bcl-2 (sc-783), anti-Bcl-xL (sc-634), anti-Mcl-1(sc-819), anti-cIAP2 (sc-7944), and anti-Cbl (sc-170) were purchased from Santa Cruz Biotechnology (Santa Cruz, California, USA). Anti-XIAP (610762) was purchased from BD Biosciences (Bedford, MA). Anti-pro-caspase-3 (ADI-AAP-113) antibody was obtained from Enzo life science (Farmington, NY). Anti-PARP (#9542) antibody, anti-cleaved caspase-3 (#9661S), anti-DR5 (#8074S), anti-cIAP1 (#4952S), anti-PSMA5 (#2457), and anti-PSMD/S5a (#1244) antibody were obtained from Cell Signaling Technology (Beverly, MA). Anti-actin (A5441) antibody was obtained from Sigma (St. Louis, MO). Other reagents were purchased from Sigma Chemical Co.

Park E.J., Min K.J., Choi K.S., Kubatka P., Kruzliak P., Kim D.E, & Kwon T.K. (2016). Chloroquine enhances TRAIL-mediated apoptosis through up-regulation of DR5 by stabilization of mRNA and protein in cancer cells. Scientific Reports, 6, 22921.

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Characterization of Cancer Cell Lines

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Human head and neck cancer AMC-HN4 cells were obtained from Asan Medical Center. MDA-MB-231, U87MG, and EA.hy926 cells were purchased from the American Type Culture Collection (Manassas, VA, USA). Primary cultured human mesangial cells (Cryo NHMC) were purchased from Clonetics (San Diego, CA, USA). The cells were cultured in Dulbecco's modified Eagle's medium that contained 10% fetal bovine serum, 20 mM Hepes buffer, and 100 μg/ml gentamicin. The PCR primers were purchased from Macrogen, Inc. (Seoul, Korea), and the other chemicals were purchased from Sigma (St. Louis, MO, USA). NAC and Trolox were obtained from Calbiochem (San Diego, CA, USA). The anti-Bcl-2, anti-Bcl-xL, anti-Mcl-1, anti-XIAP, anti-Nrf2, and anti-PARP antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The anti-cleaved caspase-3 and anti-cIAP1 antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA). The anti-pro-caspase-3 and anti-c-FLIP antibodies was obtained from ALEXIS Corporation (San Diego, CA, USA). The anti-PSMA5 antibody was purchased from Cell Signaling Technology. The anti-peroxiredoxin-SO₃ antibody was purchased from AbFRONTIER (Seoul, Korea). The anti-actin antibody was obtained from Sigma. The human Mcl-1 and c-FLIP expression vectors were constructed as described previously.^{47 (link), 48 (link)}

Seo S.U., Cho H.K., Min K.J., Woo S.M., Kim S., Park J.W., Kim S.H., Choi Y.H., Keum Y.S., Hyun J.W., Park H.H., Lee S.H., Kim D.E, & Kwon T.K. (2017). Thioridazine enhances sensitivity to carboplatin in human head and neck cancer cells through downregulation of c-FLIP and Mcl-1 expression. Cell Death & Disease, 8(2), e2599-.

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Diverse Cell Lines and Reagents

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American Type Culture Collection supplied all human cancer cells (renal carcinoma: Caki, ACHN, and A498, hepatocellular carcinoma: SK-hep1 and Huh7, breast carcinoma: MDA-MB-231) and the mouse kidney cells (TCMK-1) (Manassas, VA, USA). Primary culture of human mesangial cells (Cryo NHMC) were purchased from Clonetics (San Diego, CA, USA). Cells were grown in DMEM supplemented with 10% FBS and 100 µg/mL gentamycin. R&D system supplied z-VAD-fmk and TNF-α (Minneapolis, MN, USA), and Calbiochem supplied necrostatin-1, N-acetylcysteine (NAC) and trolox (San Diego, CA, USA). Santa Cruz Biotechnology supplied Anti-AIF antibody (Dallas, TX, USA), and Cell Signaling Technology supplied anti-PARP antibody (Beverly, MA, USA). Selleckchem supplied RLS3 (Houston, TX, USA), and BD Biosciences supplied anti-RIP1 antibody (San Jose, CA, USA). Sigma Chemical Co. supplied anti-actin antibody, Corosolic acid, cycloheximide (CHX), deferoxamine (DFO), ferrostatin-1, and α-tocopherol (St. Louis, MO, USA).

Woo S.M., Seo S.U., Min K.J., Im S.S., Nam J.O., Chang J.S., Kim S., Park J.W, & Kwon T.K. (2018). Corosolic Acid Induces Non-Apoptotic Cell Death through Generation of Lipid Reactive Oxygen Species Production in Human Renal Carcinoma Caki Cells. International Journal of Molecular Sciences, 19(5), 1309.

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Culturing Cancer Cell Lines

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Human glioma U87, U343, and T98G, human prostate carcinoma LNCaP cells, human breast carcinoma MCF-7 cells, human liver carcinoma HepG2 cells and human cervical carcinoma HeLa cells were purchased from American Tissue Type Culture Collection (Manassas, VA, USA). Cells were cultured in DMEM and RPMI 1640 medium (Invitrogen, Carlsbad, CA, USA) with 10% fetal bovine serum (HyClone, Logan, UT, USA), 1 mM L-glutamine, and 26 mM sodium bicarbonate for monolayer cell culture. Primary cultures of human astrocyte cells (Cryo NHMC) and their corresponding growth medium (CC-3146 MsGM) were purchased from Clonetics (San Diego, California, USA). p53-containing (p53^+/+) and p53-deficient (p53^−/−) HCT116 human colon carcinoma cell lines were kindly provided by Dr. Bert Vogelstein (Johns Hopkins University, Baltimore, MD, USA). These cell lines were cultured in McCoy's 5A medium (Gibco-BRL, Gaithersburg, MD, USA) containing 10% fetal bovine serum and antibiotics. The dishes containing cells were kept in a 37 °C humidified incubator with 5% CO₂.

Lee D.H., Kim D.W., Jung C.H., Lee Y.J, & Park D. (2014). Gingerol sensitizes TRAIL-induced apoptotic cell death of glioblastoma cells. Toxicology and applied pharmacology, 279(3), 253-265.

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