Bovine insulin

Manufactured by Thermo Fisher Scientific

Sourced in United States, Canada, United Kingdom, France

Bovine insulin is a lab equipment product used for research and development purposes. It is a naturally occurring hormone extracted from the pancreas of cattle, which plays a crucial role in regulating blood sugar levels. Bovine insulin can be used in various scientific applications, such as cell culture studies and pharmaceutical research.

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Insulin (904 citations)

62 protocols using bovine insulin

Osteogenic and Adipogenic Differentiation of DMSCs

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Configuration of osteogenic cells induction medium: high glucose DMEM/F12 (Genom Biotechnology, China), 2 mM glutamine (Genom Biotechnology, China), 100 nM dexamethasone (Sigma, Germany), 0.2 mm ascorbic acid (Sigma, Germany), 10 mM to-sodium glycerophosphate (Sigma, Germany). Cells were cultured in an osteogenic cells induction medium for 20 days. Osteogenic differentiation of DMSCs was assessed using alizarin red staining (Merrybaugh, China).
Configuration of Adipogenic Cells induction medium: solution A: high glucose DMEM/F12 (Genom Biotechnology, China), 2 mM glutamine (Genom Biotechnology, China), one mol/L dexamethasone (Sigma, Germany), 10 g/ml bovine insulin (GIBCO, USA), 0.5 mM IBMX (Sigma, Germany), 200 M Indomethacin (Sigma, Germany). Solution B: high glucose DMEM/F12 (Genom Biotechnology, China), 1 M dexamethasone (Sigma, Germany), 10 g/ml bovine insulin (GIBCO, USA). After three days in culture in solution A, the induction medium was replaced with Solution B, then culture for 20 days. Adipogenic differentiation was assessed using oil red O stain (Sigma, Germany).

He Y.B., Zhang L., Zhou L.L., Chen Y.M., Lu J.H., Chen J, & Liu Y.L. (2022). Effect of human follicle-stimulating hormone on immunomodulatory function of decidual mesenchymal stem cells by reducing interleukin-6 levels. Journal of Ovarian Research, 15, 60.

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Culturing and Handling Tumor Cell Lines

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Human breast adenocarcinoma MCF7 (ATCC #HTB-22), colon adenocarcinoma COLO 205 (ATCC #CCL-222), lung adenocarcinoma A549 (ATCC #CCL-185) and breast carcinoma Hs 578T (ATCC #HTB-126) cell lines were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA). COLO 205 cells were grown in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% PenStrep (PS), all purchased from Invitrogen (Grand Island, NY, USA). MCF7 cells were cultured in Eagle's minimum essential medium supplemented with 0.01 mg/mL bovine insulin, 10% FBS, and 1% PenStrep, all purchased from Invitrogen. A549 cells were grown in F-12K medium supplemented with 10% FBS, and 1% PenStrep, all purchased from Invitrogen. MCF7 cells were cultured in Dulbecco's modified Eagle's medium supplemented with 0.01 mg/mL bovine insulin, 10% FBS, and 1% PenStrep, all purchased from Invitrogen. Cell lines were incubated at 37°C and 5% CO₂ under humidified conditions, and did not exceed 90% confluence. For capture assays, tumor cells were removed from culture via treatment with trypsin-EDTA (Invitrogen) for 10 min prior to handling. All cells were washed in HBSS, and resuspended at a concentration of 1.0 × 10⁶ cells/mL in HBSS flow buffer supplemented with 0.5% HSA, 2 mM Ca²⁺, and 10 mM HEPES (Invitrogen), buffered to pH 7.4.

Mitchell M.J., Castellanos C.A, & King M.R. (2015). Immobilized Surfactant-Nanotube Complexes Support Selectin-Mediated Capture of Viable Circulating Tumor Cells in the Absence of Capture Antibodies. Journal of biomedical materials research. Part A, 103(10), 3407-3418.

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Culturing Breast Cancer Cell Lines

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MDA-MB-231, MCF-7 and MCF-10-2A cell lines were obtained from the American Type Culture Collection (ATCC). MDA-MB-231 were maintained in Leibovitz’s L15 medium (Life Technologies) supplemented with 10% FBS and antibiotic-antimytotic. MCF-7 cells were maintained in Eagle’s Minimum Essential Medium (EMEM, Life Technologies) supplemented with 10% FBS, antibiotic-antimytotic and 0.01 mg/ml bovine insulin (Life Technologies). MCF-10-2A cells were maintained in Dulbeccos modified Eagle medium (DMEM)/Ham’s F12 medium (Life Technologies) with 10% horse serum, 0.01 mg/ml bovine insulin, 20 ng/ml epidermal growth factor, 100 ng/ml Cholera toxin, 500 ng/ml hydrocortisone and antibiotic-antimytotic. Cells were incubated at 37 °C, MCF-7 and MCF-10-2A cells in the presence of 5% CO₂.

Gibbons J.A., Kanwar J.R, & Kanwar R.K. (2015). Iron-free and iron-saturated bovine lactoferrin inhibit survivin expression and differentially modulate apoptosis in breast cancer. BMC Cancer, 15, 425.

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Murine Prostate Cancer Cell Culture Protocol

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Throughout the study a murine prostate cancer cell line (transgenic adenocarcinoma of mouse prostate – TRAMP-C2) was used. TRAMP-C2 was purchased from the American Type Culture Collection and maintained in Dulbecco's modified Eagle's medium (DMEM) (Invitrogen Corp., Carlsbad, CA, USA) with 4 mM L-glutamine (Invitrogen Corp., Carlsbad, CA, USA) adjusted to contain 1.5 g/l sodium bicarbonate (Life Technologies) and 4.5 g/l glucose (Life Technologies) supplemented with 0.005 mg/ml bovine insulin (Gibco) and 5% fetal bovine serum, 5% NuSerum (Becton Dickinson), 10 nM dehydroisoandrosterone (Life Technologies) and 5 µg/ml insulin (Life Technologies).
Two hundred ninety three (human embryonic kidney) cell line was used in in vitro experiments.
Cells were cultured on 78 cm² culture plates at 37°C in a fully humidified atmosphere of 5% CO₂/95% air and passaged every 2–3 days.

Mackiewicz J., Kotlarski M., Dondajewska E., Nowicka-Kotlarska A., Krokowicz Ł, & Kazimierczak U. (2015). Cell-based Hyper-interleukin 6 or Hyper-interleukin 11 secreting vaccines combined with low dose cyclophosphamide in an orthotopic murine prostate cancer model. Contemporary Oncology, 19(3), 187-194.

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Establishing a Breast Cancer Cell Line Panel

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The BC cell lines, T-47D, MCF7, MDA-MB-231, SK-BR-3 and BT-474, and human normal mammary cells (MCF10A) were purchased from the American Type Culture Collection (ATCC). T-47D cells was maintained in RPMI-1640 (cat. no. 30-2001; ATCC) supplemented with 0.2 U/ml bovine insulin and 10% fetal bovine serum (FBS) (Gibco; Thermo Fisher Scientific, Inc.). MCF7 and MDA-MB-231 cells were maintained in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS. SK-BR-3 cells were maintained in McCoy's 5A medium (Gibco; Thermo Fisher Scientific, Inc.). BT-474 cells were maintained in ATCC Hybri-Care Medium (cat. no. 46-X) supplemented with 10% FBS. MF10A cells were maintained in RPMI-1640 supplemented with 10% FBS. All cells were cultured in an incubator at 37°C with 5% CO₂.

Kong S., Liu J., Zhang B., Lv F., Yu Y, & Qin T. (2021). MicroRNA-337-3p impedes breast cancer progression by targeting cyclin-dependent kinase 1. Oncology Letters, 23(1), 15.

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Bifidobacteria Modulation of EPEC-Induced Responses

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Rat IEC-18 cells [Catalog No. CRL-1589; American Tissue Culture Collection (ATCC), Manassas, VA, United States] were purchased from the ATCC and cultured in 95% Dulbecco’s modified Eagle’s medium supplemented with 5% fetal bovine serum and 0.1 U/mL bovine insulin (Gibco, Carlsbad, CA, United States). Bifidobacteria infantis (B. infantis), B. longum, B. bifidum and B. youth were provided by the Institute of Light Industry (Wuhan, China). EPEC (serotype O127: B8) endotoxin was purchased from Sigma-Aldrich (St Louis, MO, United States) and constituted for use at a concentration of 1 mg/mL.
Six groups were established in this experiment, including normal control, EPEC, B. infantis, B. longum, B. bifidum, and B. youth. The EPEC group received intervention with the EPEC endotoxin, whereas the bifidobacteria groups underwent treatment with B. infantis, B. longum, B. bifidum and B. youth separately. Normal controls did not undergo any intervention.

Yang X., Gao X.C., Liu J, & Ren H.Y. (2017). Effect of EPEC endotoxin and bifidobacteria on intestinal barrier function through modulation of toll-like receptor 2 and toll-like receptor 4 expression in intestinal epithelial cell-18. World Journal of Gastroenterology, 23(26), 4744-4751.

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Culturing Normal Human Corneal Epithelial Cells

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Normal human corneal epithelial cell line 10.014 pRSV-T (ATCC No. CRL-11515) was used for the experiments. The cells were cultured as monolayers in 25 cm² culture flasks (Nunc. Roskilde, Denmark) coated with PureCol ™ ultrapure collagen (INAMED Biomaterials, Fremont, CA, USA) at 3.1 mg/ml concentration. Cell lines were maintained in defined keratinocyte-serum free medium (K-SFM) (Gibco, Gibco ™, Paisley, UK) supplemented with 75 µg/ml endothelial cell growth factor (ECGF) (Sigma-Aldrich Co., St. Louis, MO, USA), 0.05 mg/ml bovine pituitary extract (BPE) (Gibco, Gibco ™, Paisley, UK), 500 ng/ml hydrocortisone (Sigma-Aldrich Co., St. Louis, MO, USA) and 0.0005 mg/ml bovine insulin (Gibco, Gibco ™, Paisley, UK) and antibiotics (100 U/ml penicillin, 100 µg/ml streptomycin) (Sigma-Aldrich Co., St. Louis, MO, USA) at 37°C in a humidified atmosphere with 5% CO₂.

Paduch R, & Woźniak A. (2015). The Effect of Lamium album Extract on Cultivated Human Corneal Epithelial Cells (10.014 pRSV-T). Journal of Ophthalmic & Vision Research, 10(3), 229-237.

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Wound Healing Scratch Assay for Cell Migration

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Cells were assessed in wound healing scratch assays using the IncuCyte (Essen Bioscience). Cell migration assays were achieved by growing MCF7 and MDA-MB-231 cells to confluence in 96-well plates (Essen BioScience; cat. no. 4379). Wounds were made using the 96-pin wound-making tool (WoundMaker; Essen Bioscience) to simultaneously create a precise and reproducible wound in each well, 1 h after the plate was washed twice with PBS, and incubated with DMEM containing 1% FBS, 1% penicillin-streptomycin (Gibco), 4 mM L-glutamine (Gibco), sodium pyruvate (1 mM) and 0.01 mg/mL bovine insulin (Sigma). Cell migration was monitored in real time by IncuCyte, and wound width was measured by the IncuCyte software Zoom (version 2014a, Essen Bioscience, Ann Arbor, MI, USA).

Mahadevappa R., Neves H., Yuen S.M., Jameel M., Bai Y., Yuen H.F., Zhang S.D., Zhu Y., Lin Y, & Kwok H.F. (2018). DNA Replication Licensing Protein MCM10 Promotes Tumor Progression and Is a Novel Prognostic Biomarker and Potential Therapeutic Target in Breast Cancer. Cancers, 10(9), 282.

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Cultivation of Rat Intestinal and Canine Kidney Cells

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The rat intestinal epithelial cell line IEC-6 was obtained from the American Type Culture Collection (Manassas, VA). Cells were cultured in Dulbecco’s modified Eagle’s medium (Sigma-Aldrich) containing 4.5 g/l L-glucose, 3.7 g/l sodium bicarbonate, and 4 mM L-glutamine and supplemented with 0.1 U/ml bovine insulin and 10 % heat-inactivated fetal bovine serum (Gibco, Langley, OK) ^{2 (link), 29 (link)}. Madin-Darby canine kidney cells (MDCK) stably expressing G93T hNIS were cultured as described ^{13 (link)}.

Nicola J.P., Carrasco N, & Amzel L.M. (2014). Physiological sodium concentrations enhance the iodide affinity of the Na+/I− symporter. Nature communications, 5, 3948.

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Cultivation of Rat Intestinal and Canine Kidney Cells

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Nicola J.P., Carrasco N, & Amzel L.M. (2014). Physiological sodium concentrations enhance the iodide affinity of the Na+/I− symporter. Nature communications, 5, 3948.

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