Nuserum 4

Manufactured by BD

Sourced in United States

NuSerum IV is a laboratory instrument designed for automated serum and plasma separation. It utilizes a centrifugation process to separate biological samples into distinct components, such as serum or plasma, for further analysis.

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Lab products found in correlation

Human insulin, by Merck Group (3 mentions) Dmem medium, by Merck Group (2 mentions) Media 254, by Thermo Fisher Scientific (2 mentions) Rat tail collagen 1, by BD (2 mentions) Fetal bovine serum (fbs), by Harvard Bioscience (2 mentions) Sodium pyruvate, by Harvard Bioscience (2 mentions) Non essential amino acids (neaa), by Harvard Bioscience (2 mentions) Rpmi 1640 medium, by Merck Group (2 mentions) Antibiotic antimycotic solution, by Merck Group (2 mentions) L glutamine, by Harvard Bioscience (2 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Streptomycin sulphate, by Thermo Fisher Scientific (1 mentions) Insulin, by Merck Group (1 mentions) Tramp c2, by American Type Culture Collection (1 mentions) Dihydrotestosterone, by Merck Group (1 mentions) C3h mice, by Taconic Biosciences (1 mentions) Annexin 5 fitc detection kit, by BD (1 mentions) Dexamethasone (dex), by Merck Group (1 mentions) Hepatocyte growth factor (hgf), by R&D Systems (1 mentions)

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NuSerum (83 citations)

9 protocols using nuserum 4

Cell Line Cultivation Protocols

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All cell lines used in the study were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Human breast adenocarcinoma MDA-MB-231, prostate carcinoma DU-145, cervix carcinoma HeLa, osteosarcoma U2-OS, glioblastoma U373, U-87 MG and T98-G, and telomerase-immortalised retinal epithelium RPE-1-hTERT cell lines were cultured in high-glucose (4.5 g/l) DMEM. Media were supplemented with 10% foetal bovine serum (Gibco/Thermo Fisher Scientific, Waltham, MA), 100 U/ml penicillin, and 100 μg/ml streptomycin sulphate (Gibco/Thermo Fisher Scientific, Waltham, MA). Cells were kept at 37 °C under 5% CO₂ atmosphere and 95% humidity.
The mouse TC-1 cell line was obtained in vitro by co-transfection of murine lung C57BL/6 cells with HPV16 E6/E7 and activated human H-Ras (G12V) oncogenes. TC-1 cells were cultured in RPMI-1640 medium supplemented with 10% foetal calf serum, l-glutamine, and antibiotics³⁵ (link). Prostate carcinoma cell line TRAMP-C2³⁶ (link) was maintained in DMEM medium (Sigma-Aldrich, St. Louis, MO) supplemented with 5% FCS, Nu-Serum IV (5%; BD Biosciences, Bedford, MA), 5 µg/ml human insulin (Sigma-Aldrich, St. Louis, MO), dehydroisoandrosterone (DHEA, 10 nM; Sigma-Aldrich, St. Louis, MO), and antibiotics³⁷ (link).

Oleksak P., Psotka M., Vancurova M., Sapega O., Bieblova J., Reinis M., Rysanek D., Mikyskova R., Chalupova K., Malinak D., Svobodova J., Andrys R., Rehulkova H., Skopek V., Ngoc Lam P., Bartek J., Hodny Z, & Musilek K. (2021). Design, synthesis, and in vitro evaluation of BP-1-102 analogs with modified hydrophobic fragments for STAT3 inhibition. Journal of Enzyme Inhibition and Medicinal Chemistry, 36(1), 410-424.

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Transgenic Cell Lines for Immunotherapy

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TC-1 cell line was obtained by in vitro co-transfection of murine lung C57BL/6 cells with HPV16 E6/E7 and activated human H-Ras (G12V) oncogenes [33 (link)]. IL-12-gene modified TC-1/IL-12 (231/clone 15) cells used for immunotherapy produced in vitro 40 ng IL-12/1 × 10⁵ cells/ml medium/48 h and were irradiated (150 Gy) before use [34 (link)]. TC-1 cells were cultured in RPMI 1640 medium supplemented with 10% fetal calf serum, L-glutamine and antibiotics. TRAMP-C2 prostate carcinoma cells [35 (link)] were obtained from ATCC collection. TRAMP-C2 cells were maintained in D-MEM medium (Sigma-Aldrich, Saint Louis, MO, USA) supplemented with 5% FCS, Nu-Serum IV (5%; BD Biosciences, Bedford, MA, USA), 5 mg/ml human insulin (Sigma-Aldrich), dehydroisoandrosterone (DHEA, 10 nM; Sigma-Aldrich) and antibiotics [36 (link)].

Simova J., Sapega O., Imrichova T., Stepanek I., Kyjacova L., Mikyskova R., Indrova M., Bieblova J., Bubenik J., Bartek J., Hodny Z, & Reinis M. (2016). Tumor growth accelerated by chemotherapy-induced senescent cells is suppressed by treatment with IL-12 producing cellular vaccines. Oncotarget, 7(34), 54952-54964.

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TRAMP-C2 Prostate Tumor Implantation

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Specific pathogen free C57BL/6 mice and C3H mice, 6 to 8 weeks of age, were purchased from Taconic Farms (Germantown, NY). TRAMP-C2 (ATCC CRL-2731; originally derived from the prostate tumor of a TRAMP mouse on the C57BL/6 background) cells were used for tumor challenge studies. TRAMP-C2 cells were grown and expanded in vitro with IMDM medium supplemented with 5% Fetal bovine serum (FBS; Gemini, Sacramento, CA), 5% Nu Serum IV (BD Biosciences, San Jose, CA), 0.01 nM dihydrotestosterone (Sigma Chemical Co.), and 5 μg/ml insulin (Sigma Chemical Co.). All in vivo studies were in compliance and approved by University of Southern California Institutional Animal Care and Use Committee (USC IACUC).

Yan L., Da Silva D.M., Verma B., Gray A., Brand H.E., Skeate J.G., Porras T.B., Kanodia S, & Kast W.M. (2014). Forced LIGHT expression in prostate tumors overcomes Treg mediated immunosuppression and synergizes with a prostate tumor therapeutic vaccine by recruiting effector T lymphocytes. The Prostate, 75(3), 280-291.

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Isolation and Apoptosis Analysis of Intrahepatic BECs

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To isolate the intrahepatic BECs, a two-step liver perfusion was performed. The perfused liver was removed and hepatocytes were selectively removed by forcing them with gentle pressure through an incision in the liver. Cells were suspended in DMEM media supplemented with 10% FBS, 5% NuSerum IV (BD), 0.5 mg/ml insulin–transferrin–sodium selenite (Gibco), 1 mmol/l ascorbic acid 2-phosphate, 10K7 M dexamethasone (Sigma-Aldrich Corp.), 10 ng/ml EGF (R&D, Minneapolis, MN, USA), 10 ng/ml HGF (R&D, Minneapolis, MN, USA). After 10 days cells were exposed to ionomycine 1 μg/ml for 22 hours and percentage of apoptotic cells was determined by flow cytometry using the Annexin V FITC Detection Kit (BD Pharmingen, San Jose, CA).

Arsenijevic A., Milovanovic M., Milovanovic J., Stojanovic B., Zdravkovic N., Leung P.S., Liu F.T., Gershwin M.E, & Lukic M.L. (2016). Deletion of Galectin-3 Enhances Xenobiotic Induced Murine Primary Biliary Cholangitis by Facilitating Apoptosis of BECs and Release of Autoantigens. Scientific Reports, 6, 23348.

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Diverse Melanoma and Endothelial Cell Lines

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B16F0, B16F1, B16F10 (ATCC), ΔBraf/Pten−/− (derived from B6.Cg-Braf^tm1MmcmPten^tm1HwuTg^{(Tyr-cre/ERT2)}13Bos/BosJ mice (UNC-Chapel Hill Mouse Phase 1 Unit)) cell lines PBT2460 and PBT2130 (Drs. W. Kim and J. Bear, UNC-Chapel Hill), and human melanoma cell lines WM1158, SBCl2, Sk-Mel-119, Sk-Mel-173, and WM2664 (Dr. J. Shields, UNC-Chapel Hill) were maintained in DMEM with 4.5 g/mL D-Glucose and 10% FBS. Human melanoma cell lines RPMI7951, RPMI8322, and Mel505 (Dr. J Shields, UNC-Chapel Hill) were maintained in RPMI with 10% FBS. The normal human melanocyte line NHM7 (Dr. J Shields, UNC-Chapel Hill) was maintained in Media 254 (Gibco) supplemented with HGMS (Gibco). EC (isolated by us previously ^{8 (link)}) were maintained in Endothelial Cell Media (EC-Media), composed of DMEM with 1g/L glucose, 5 μg/L bFGF, 10 μg/L VEGF, 100 mg/L heparin, antibiotics, and 10% NuSerum IV (BD).

Dunleavey J.M., Xiao L., Thompson J., Kim M.M., Shields J.M., Shelton S.E., Irvin D.M., Brings V.E., Ollila D., Brekken R.A., Dayton P.A., Melero-Martin J.M, & Dudley A.C. (2014). Vascular channels formed by subpopulations of PECAM1+ melanoma cells. Nature communications, 5, 5200.

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In vitro Model of the Blood-Brain Barrier

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Human brain microvascular endothelial cell (HBMEC) line was used as a simplified and validated in vitro model of the BBB^{32 (link), 44 (link), 45 (link), 62}. This cell line was derived from primary cultures of HBMEC transfected with SV40 large T antigen^{63 (link)}. HBMEC line was cultured in RPMI 1640 medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS - Biochrom AG), 10% NuSerum IV (BD Biosciences), 1% non-essential amino acids (NEAA - Biochrom AG), 1% minimal essential medium (MEM) vitamins (Biochrom AG), 1 mM sodium pyruvate (Biochrom AG), 2mM L-glutamine (Biochrom AG), and 1% antibiotic-antimycotic solution (Sigma-Aldrich). For immunostaining and cytoprotection studies, cells were seeded at a density of 8 × 10⁴ cell/mL in 24-well and 96-well plates, respectively, and treated after 48 h in culture. For integrity and transport studies, cells were seeded on polyester transwell inserts (0.4 μm, Corning Costar Corp., USA) at a density of 8 × 10⁴ cell/insert and treated after 8 days in culture. Inserts and plates were coated with rat-tail collagen-I (BD Biosciences, Erembodegem, Belgium) before seeding. All experiments were performed after monolayer formation.

Figueira I., Garcia G., Pimpão R.C., Terrasso A.P., Costa I., Almeida A.F., Tavares L., Pais T.F., Pinto P., Ventura M.R., Filipe A., McDougall G.J., Stewart D., Kim K.S., Palmela I., Brites D., Brito M.A., Brito C, & Santos C.N. (2017). Polyphenols journey through blood-brain barrier towards neuronal protection. Scientific Reports, 7, 11456.

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Diverse Melanoma and Endothelial Cell Lines

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Development of Murine Tumor Cell Lines

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The TC-1 tumor cell line (obtained from the ATCC collection) was developed by co-transfection of murine C57BL/6 lung cells with HPV16 E6/E7 genes and activated (G12V) Ha-ras plasmid DNA (25 (link)). TRAMP-C2 tumor cells (obtained from the ATCC collection), MHC class I-deficient, were established from a heterogeneous 32-week tumor of the transgenic adenocarcinoma mouse prostate (TRAMP) model (26 (link)). TC-1 cells were maintained in RPMI-1640 medium (Sigma-Aldrich GmbH, Steinheim, Germany) supplemented with 10% FCS (PAN Biotech GmbH, Aidenbach, Germany), 2 mM L-glutamine and antibiotics; TRAMP-C2 cells were maintained in D-MEM medium (Sigma-Aldrich) supplemented with 5% FCS, Nu-Serum IV (5%; BD Biosciences, Bedford, MA, USA), 0.005 mg/ml human insulin (Sigma-Aldrich), dehydroisoandrosterone (DHEA, 10 nM; Sigma-Aldrich) and antibiotics. Both cell lines were cultured at 37ºC in a humidified atmosphere with 5% CO₂ cells. In the in vivo experiments, 5×10⁴ TC-1 cells and 1×10⁶ TRAMP-C2 cells were administered for the challenge. In our hands, 5×10⁴ TC-1 cells represent 5 TID₅₀ doses and 1×10⁶ TRAMP-C2 cells represent 3 TID₅₀ doses.

MIKYŠKOVÁ R., ŠTĚPÁNEK I., INDROVÁ M., BIEBLOVÁ J., ŠÍMOVÁ J., TRUXOVÁ I., MOSEROVÁ I., FUČÍKOVÁ J., BARTŮŇKOVÁ J., ŠPÍŠEK R, & REINIŠ M. (2015). Dendritic cells pulsed with tumor cells killed by high hydrostatic pressure induce strong immune responses and display therapeutic effects both in murine TC-1 and TRAMP-C2 tumors when combined with docetaxel chemotherapy. International Journal of Oncology, 48(3), 953-964.

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Established Biochemical Assay Protocols

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The basal medium Roswell Park Memorial Institute 1640, antibiotic-antimycotic solution, human serum albumin (fraction V, fatty acid free), bovine serum albumin, Hoechst 33258 dye, sodium fluorescein and UCB were purchased from Sigma Chemical Co. (St. louis, MO, USA). Non-essential amino acids, sodium pyruvate, l-glutamine, fetal bovine serum and minimum essential medium vitamins were from Biochrom AG (Berlin, Germany). Nuserum IV and rat-tail collagen I were acquired from BD Biosciences (Erembodegem, Belgium). TRIzol Plus RNA Purification Kit, was from Invitrogen (Carlsbad, CA, USA). Caspase-3 substrate and Ac-Asp-Glu-Val-Asp-p-nitroanilide, were acquired from Calbiochem (San Diego, CA, USA). DuoSet ELISA kit was from R&D systems (Minneapolis, MN, USA). Primers for real-time PCR analysis were purchased from Thermo Scientific (Soeborg, Denmark). RevertAid H Minus First Strand cDNA synthesis and Maxima SYBR Green qPCR Master Mix (2×) were obtained from Fermentas (Burlington, ON, Canada). All other chemicals were of analytical grade and were purchased from Merck (Darmstadt, Germany).

Palmela I., Correia L., Silva R.F., Sasaki H., Kim K.S., Brites D, & Brito M.A. (2015). Hydrophilic bile acids protect human blood-brain barrier endothelial cells from disruption by unconjugated bilirubin: an in vitro study. Frontiers in Neuroscience, 9, 80.

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