10 cm dishes

Manufactured by BD

Sourced in United States

The 10-cm dishes are laboratory equipment used for various applications in cell culture and microbiology. They provide a standard-sized petri dish for growing and maintaining cell cultures or microorganisms. The dishes are made of high-quality materials and designed to provide a sterile, controlled environment for experimental purposes.

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Lab products found in correlation

Tris glycine gel, by Bio-Rad (2 mentions) Alexa fluor 800 rabbit anti mouse, by Thermo Fisher Scientific (2 mentions) Alexafluor 680 goat anti rabbit, by Thermo Fisher Scientific (2 mentions) Odyssey infrared imaging system, by LI COR (2 mentions) Ripa buffer, by Cell Signaling Technology (2 mentions) β actin, by Santa Cruz Biotechnology (2 mentions) Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions) Collagen coated, by Merck Group (1 mentions) Transit lenti reagent, by Mirus Bio (1 mentions) 24 well plate, by BD (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions) Dithiothreitol (dtt), by Merck Group (1 mentions) Dmem f12, by Thermo Fisher Scientific (1 mentions) Sodium dodecyl sulfate (sds), by Merck Group (1 mentions) Fetal bovine serum (fbs), by Biosera (1 mentions) Glycerol, by BD (1 mentions) Lb broth, by Merck Group (1 mentions) Penicillin streptomycin, by Lonza (1 mentions) Dulbecco modified eagle medium (dmem), by PAN Biotech (1 mentions)

10 protocols using 10 cm dishes

Duxbl Knockdown with shRNA Vectors

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For Duxbl knockdown, MSCV-LMP empty vector plus four different MSCV-LMP vectors containing Duxbl-directed shRNAs were generated using the following oligo templates: sh2-1, 5′-TGCTGTTGACAGTGAGCGAACTCTTCGTGTGTGGTTTGAATAGTGAAGCCACAGATGTATTCAAACCACACACGAAGAGTGTGCCTACTGCCTCGGA-3′; sh2-2, 5′-TGCTGTTGACAGTGAGCGCAGAATCCCAATCCTGATCTAGTAGTGAAGCCACAGATGTACTAGATCAGGATTGGGATTCTTTGCCTACTGCCTCGGA-3′; sh3-1, 5′-TGCTGTTGACAGTGAGCGCTTGCATGCTGTTCTGAAGAAATAGTGAAGCCACAGATGTATTTCTTCAGAACAGCATGCAAATGCCTACTGCCTCGGA-3′; and sh3-2, 5′-TGCTGTTGACAGTGAGCGAAGTTTGCATGCTGTTCTGAAGTAGTGAAGCCACAGATGTACTTCAGAACAGCATGCAAACTCTGCCTACTGCCTCGGA-3′.
A total of 5 × 10⁶ Plat-E cells were plated 1 d before transfection in 10-cm dishes (Falcon) with 10 ml DMEM (Sigma) supplemented with 10% FBS, 100 U/ml penicillin (BioConcept), 100 mg/ml streptomycin (BioConcept), and 1 mM sodium pyruvate (Sigma). The next day, 20 µg plasmid and 80 µg polyethylenimine were mixed in 1 ml DMEM without supplements, incubated for 15 min at room temperature, and subsequently added dropwise to the Plat-E cells. 24 h after transfection, supernatant was replaced with fresh medium. Viral supernatants were collected 48 and 72 h after transfection and either used directly for transductions or stored at −80°C.

Klein F., Mitrovic M., Roux J., Engdahl C., von Muenchow L., Alberti-Servera L., Fehling H.J., Pelczar P., Rolink A, & Tsapogas P. (2019). The transcription factor Duxbl mediates elimination of pre-T cells that fail β-selection. The Journal of Experimental Medicine, 216(3), 638-655.

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Protein Expression Analysis in RAW 264.7 Cells

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RAW 264.7 cells (1×107 cells) were cultured in 10-cm dishes (Falcon) and allowed to adhere for 24 h. After treatment with FA 100μM 1 followed by co-incubation with LPS 100 ngr/ml for 4 h, the cells were washed twice with cold PBS. Whole-cell lysates were obtained using RIPA buffer (Cell Signaling Inc. Beverly, MA, USA). The protein concentration of cell lysates was determined by the Bradford method. An amount of protein (30 μg) was separated on 8–16% Tris-Glycine Gel (BioRad) gels by electrophoresis and transferred to a nitrocellulose membrane. The membranes were subsequently incubated for 1 h at room temperature with 3% BSA in TBS buffer (0.1% v/v) to block non-specific binding and incubated with an appropriate primary antibody in 1% BSA in TBST (tween 0.01% v/v). Antibodies polyclonal anti-mouse recognizing HSP90, lamin B1, Keap1, COX2, and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Incubation with the secondary antibodies Alexa Fluor 680 goat anti-rabbit (1:2000) and Alexa Fluor 800 rabbit anti-mouse (1:5000) (Molecular Probes, Life Technologies, Carlsbad, CA, USA) was performed for 1 h at room temperature. Densitometry analysis was conducted using the Odyssey Infrared Imaging System (Li-COR Bioscience, NE, USA).

Bonura A., Giacomarra M, & Montana G. (2022). The Keap1 signaling in the regulation of HSP90 pathway. Cell Stress & Chaperones, 27(3), 197-204.

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Lentiviral Particle Production in HEK293T

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Lentiviral particles were produced in HEK293T cells. On day −1, HEK293T cells were seeded in collagen-coated (Sigma-Aldrich) 10-cm dishes (Falcon) at a density of 6 × 10⁶ cells per dish. On day 0, cells were transfected with 5 µg packaging plasmid mix (pMDLg/pRRE, pRSV-Rev and pMD2.G; ratio 4:1:1) and 5 µg transfer plasmid (pLV-TetO-hNGN2-eGFP-puro, FudeltaGW-rtTA, myc-tagged SYNJ2BP WT, S21A or S21E) using TransIT-Lenti reagent (Mirus Bio). On day 2, the medium containing lentiviral particles was mixed with lentivirus precipitation solution (Alstem; ratio 4:1) and incubated at 4 °C overnight. On day 3, the lentiviral particles were collected by centrifugation at 1,500g for 30 min, resuspended in 1 ml ice-cold PBS/dish and stored at −80 °C.

Hees J.T., Wanderoy S., Lindner J., Helms M., Murali Mahadevan H, & Harbauer A.B. (2024). Insulin signalling regulates Pink1 mRNA localization via modulation of AMPK activity to support PINK1 function in neurons. Nature Metabolism, 6(3), 514-530.

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Western Blot Analysis of Inflammatory Markers

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A549 cells (1 × 10⁷ cells) were cultured in 10-cm dishes (Falcon) and allowed to adhere for 24 h. After treatment with BMS- 345,541 1 μM for 1 h, followed by incubation with LPS 100 ngr/ml for 4 h, the cells were washed twice with cold PBS. Whole-cell lysates were obtained using RIPA buffer (Cell Signaling Inc. Beverly, MA, USA). The protein concentration of cell lysates was determined by the Bradford method. An amount of protein (30 μg) was separated on 8–16% Tris–Glycine Gel (BioRad) gels by electrophoresis and transferred to a nitrocellulose membrane. The membranes were subsequently incubated for 1 h at room temperature with 3% BSA in TBS buffer (0.1% v/v) to block non-specific binding and incubated with an appropriate primary antibody in 1% BSA in TBST (tween 0.01% v/v). Antibodies polyclonal antimouse recognizing HSP90, IKKα/β, COX2, HO1 and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); antibody polyclonal antirabbit recognizing p-IKKα/β was purchased from Cell Signaling Technology Massachussetts, USA. Incubation with the secondary antibodies Alexa Fluor 680 goat anti-rabbit (1:2000) and Alexa Fluor 800 rabbit anti-mouse (1:5000) (Molecular Probes, Life Technologies, Carlsbad, CA, USA) was performed for 1 h at room temperature. Densitometry analysis was conducted using the Odyssey Infrared Imaging System (LiCOR Bioscience, NE, USA).

Giacomarra M., La Torre M, & Montana G. (2023). Effects of Inhibition of IKK Kinase Phosphorylation On the Cellular Defence System and HSP90 Activity. Inflammation, 47(1), 74-83.

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Culturing SH-SY5Y cells with terrein

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SH-SY5Y and SH-SY5Y expressing HA-tagged mAng1 (SH-SY5Y mAng1HA) were maintained in DMEM:F12 (Invitrogen) supplemented with 10% FBS (Biosera), 1% Non-essential amino acids (Invitrogen) and 1% Glutamax (Invitrogen)) on 10 cm dishes (BD Falcon). SH-SY5Y cell lines were seeded at a density of 10⁵ cells/cm² on acid-washed coverslips (SLS) in 24-well plates (BD Falcon) in complete medium. Medium was exchanged after 24 h with complete medium containing 0, 15, 20 or 30 µM terrein. Cells were cultured in the presence of terrein for the indicated period then fixed with 4% PFA on ice for 15 min. After fixing cells were washed twice with PBS and dehydrated to 70% ethanol through 30% and 50% in ten minute incubations each. For western blotting cells were lysed in a reducing and denaturing buffer containing 2% SDS (Sigma), 10% Glycerol (BDH), 60 mM Tris (Sigma) and 100 mM DTT (Sigma).

Ferguson R., Holloway D.E., Chandrasekhar A., Acharya K.R, & Subramanian V. (2019). The catalytic activity and secretion of zebrafish RNases are essential for their in vivo function in motor neurons and vasculature. Scientific Reports, 9, 1107.

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Infection Assay with A549 Cells

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We used the human cell line A549 (DMSZ, ACC 107), which closely recapitulates the AECII phenotype as our model system (Nardone and Andrews, 1979 (link); Foster et al., 1998 (link)). Cells were propagated in DMEM (PAN-Biotech) containing 10% FBS (PAN-Biotech) and 1% Penicillin/Streptomycin (Lonza). For infection experiments, cells were washed with PBS and seeded into 6-well plates (Greiner bio-one) or 10cm dishes (Falcon) in an antibiotic-free medium containing 1% FBS.
Bacteria (E. coli ATCC 25922 and K. pneumoniae ATCC 43816) were grown from overnight cultures under sterile conditions in LB Broth (Sigma-Aldrich) to late logarithmic phase [optical density 600nm (OD600) 0.45-0.6]. Bacterial counts were determined before each experiment using a cell counter and analyzer (CASY, 45µm capillary, OLS OMNI Life Science).
For growth assays, bacteria were diluted to OD₆₀₀ of 0.005 in cell culture medium in 96-well plates (Greiner bio-one), and directly afterwards incubated in an automated microplate reader (Spark, TECAN) at 37°C, 5% CO2 under constant double orbital shaking. OD₆₀₀ was measured every 5min for a total of 10h.
For heat-inactivation, bacteria were incubated at 70°C for 20min and afterwards plated on LB plates to confirm absence of viable bacteria.

Grubwieser P., Hoffmann A., Hilbe R., Seifert M., Sonnweber T., Böck N., Theurl I., Weiss G, & Nairz M. (2022). Airway Epithelial Cells Differentially Adapt Their Iron Metabolism to Infection With Klebsiella pneumoniae and Escherichia coli In Vitro. Frontiers in Cellular and Infection Microbiology, 12, 875543.

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Stable 4T1 Reporter Cell Line Development

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4T1 mammary carcinoma cells (ATCC) at 95% confluency were co-transfected with 10 µg of pκB₅:IκBα-FLuc^{52 (link)} and 3 µg of pIRES-puro plasmid DNA using Fugene 6 (Roche) in 10 cm dishes (BD Bioscience). Cultures were incubated at 37 °C and after 24 h, the media was replaced with fresh RPMI supplemented with 10% heat-inactivated FBS. 24 h later, cells were split at multiple dilutions into media containing 0.5 µg/ml puromycin to select for stable transformants. After two weeks, isolated cell colonies were imaged to confirm reporter gene expression, and bioluminescent colonies were harvested and expanded. Reporter cells were continuously cultured in the presence of 0.5 µg/ml puromycin to maintain the expression of the reporter plasmid.

Gonzalez C., Williamson S., Gammon S.T., Glazer S., Rhee J.H, & Piwnica-Worms D. (2023). TLR5 agonists enhance anti-tumor immunity and overcome resistance to immune checkpoint therapy. Communications Biology, 6, 31.

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Isolation and Culture of Endometrial Stromal Cells

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Endometrial tissue was minced and digested with phosphate-buffered saline (PBS) containing 0.3 mg/ml collagenase III (Worthington Biochemical Corporation, Freehold, NJ, United States) and 40 μg/ml deoxyribonuclease type I (Worthington Biochemical Corporation) in a shaking water bath for 60 min at 37°C (Xiang et al., 2014 (link)). The endometrial tissue underwent two rounds of digestion, and the dispersed cells were filtered through 40-μm sieves (BD Bioscience, San Jose, CA, United States). The red blood cells, cell debris, and cell clumps were removed by Ficoll-Paque (GE Healthcare, Uppsala, Sweden) density-gradient centrifugation. Anti-CD45 antibody-coated Dynabeads (Invitrogen, Waltham, MA, United States) were used for the removal of leukocytes, and anti-CD326 antibody-coated microbeads (Miltenyi Biotec Inc., San Diego, CA, United States) were used to separate epithelial cells from stromal cell population. Freshly purified stromal cells were seeded onto 10-cm dishes (BD Biosciences, San Jose, CA, United States) coated with fibronectin (1 mg/ml, Invitrogen) containing growth medium (GM) with 1% penicillin (Invitrogen), 1% L-glutamine (Invitrogen), and 10% FBS (Invitrogen) in DMEM/F-12 (Sigma-Aldrich, St Louis, MA, United States). Stromal cells were cultured for 1–2 weeks in a humidified carbon dioxide incubator at 37°C. The medium was changed every 7 days.

Li T., Chan R.W., Lee C.L., Chiu P.C., Li R.H., Ng E.H, & Yeung W.S. (2022). WNT5A Interacts With FZD5 and LRP5 to Regulate Proliferation and Self-Renewal of Endometrial Mesenchymal Stem-Like Cells. Frontiers in Cell and Developmental Biology, 10, 837827.

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Culturing Glioblastoma Cell Lines

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We used the human GBM cell lines T98G and U87MG and one murine GBM cell line, GL261. T98G cells were purchased from the JCRB Cell Bank, and U87MG cells were from the American Type Culture Collection. GL261 cells were kindly provided by Dr. Masanao Saio (Gunma University, Japan).
As previously described, 21 T98G, U87MG, and GL261 cells were cultured in Dulbecco's modified Eagle medium (DMEM) with low glucose (Nacalai Tesque), supplemented with 10% fetal bovine serum (FBS; Valeant), 100 U/mL penicillin, and 100 μg/ml streptomycin in 10-cm dishes (BD Biosciences) at 37°C in 5% CO 2 .

Yamada T., Tsuji S., Nakamura S., Egashira Y., Shimazawa M., Nakayama N., Yano H., Iwama T, & Hara H. (2021). Riluzole enhances the antitumor effects of temozolomide via suppression of MGMT expression in glioblastoma. Journal of neurosurgery, 134(3).

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Isolation and Differentiation of Murine Neutrophils

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Bone marrow cells were sterilely harvested from tibiae and femurs of C3H/HeJ mice. After lysing red blood cells, bone marrow cells were cultured at a cell density of 2×10⁶ per ml in 10-cm dishes (BD Biosciences, Durham, NC, USA) at 37°C in an atmosphere of 5% CO₂. Cells were cultured in RPMI 1640 medium supplemented with 10% fetal calf serum (cat#100-106, Gemini Bio-Products, West Sacramento, CA, USA), 2 mM l-glutamine (cat#G7513, Sigma, St. Louis, MO, USA), 1X MEM non-essential amino acids (cat#11140-050, Gibco, Grand Island, NY, USA), 1 mM sodium pyruvate (cat#25-000-CI, Cellgro, Manassas, VA, USA), 0.5 mM beta-mercaptoethanol (cat#M3148, Sigma), 100 U/ml penicillin G, and 100 μg/ml streptomycin (cat#30-002-CI, Cellgro).
To make neutrophils, bone marrow cells were cultured in growth medium containing 100 ng/ml recombinant G-CSF (cat#250-05, PeproTech). Every 3 days, floating cells were harvested and re-cultured in fresh growth medium. On day 8, cells floating in the supernatants were analyzed by flow cytometry for the presence of Ly-6G or Gr1. When positivity was >80%, the cells were used as neutrophils in chemotaxis assays.

Hou S., Xu R., Zhu C., Shan S., Han L, & Wang H. (2018). Chlamydial Plasmid-Encoded Protein pGP3 Inhibits Development of Psoriasis-Like Lesions in Mice. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 24, 5159-5167.

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