Collagen-coated glass-bottomed dishes (MatTek, Ashland, MA) were coated with 0.2% Matrigel in serum-free medium for 30 min at 37°C. Cells were seeded sparsely overnight and then serum starved for 4 h in Leibowitz’s L-15 medium (Life Technologies) with 0.35% BSA. Differential interference contrast images were acquired every 20 s for 10 min in an environmentally controlled microscope (TE2000; Nikon) with a 20× objective and a Photometrics CoolSNAP HQ camera. Growth factor/inhibitor solutions were added after 80 s. Cell areas were traced immediately before and 9 min after stimulation using ImageJ (National Institutes of Health, Bethesda, MD). Data shown are from individual cells (at least 60 overall) pooled from at least three separate experiments.
Leibowitz s l15 medium
Manufactured by Thermo Fisher Scientific
Sourced in United States
Leibowitz's L15 medium is a cell culture medium formulated for the growth and maintenance of various cell types. It is a basal medium that provides the necessary nutrients and growth factors to support cell proliferation and survival in vitro. The medium is widely used in cell biology research and applications that require the cultivation of diverse cell lines.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Te 2000e inverted microscope, by Oxford Instruments (2 mentions)
603 1.49 na tirf objective, by Oxford Instruments (2 mentions)
Xd spinning disk confocal system, by Oxford Instruments (2 mentions)
Opti mem, by Thermo Fisher Scientific (2 mentions)
Collagen coated glass bottomed dishes, by MatTek (1 mentions)
Dharmafect, by Horizon Discovery (1 mentions)
Lipofectamine, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Imatinib, by Merck Group (1 mentions)
Puromycin, by Thermo Fisher Scientific (1 mentions)
Anti p21, by Merck Group (1 mentions)
Anti p27, by Merck Group (1 mentions)
Rabbit anti gapdh, by Merck Group (1 mentions)
Tryptose phosphate broth, by Merck Group (1 mentions)
Fetal calf serum (fcs), by Lonza (1 mentions)
10 protocols using leibowitz s l15 medium
1
Cell Morphology Dynamics Imaging
2
Culturing HeLa, MCF7, and MDA-MB-231 cell lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa and MCF7 cell lines were cultured in DMEM (Life Technologies, Paisley, UK) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, Dorset, UK) and maintained at 37°C and 5% CO2. MDA-MB-231 cells were cultured in Leibowitz’s L-15 medium (Life Technologies) supplemented with 15% FBS and 2 mM l -glutamine (Life Technologies) and maintained at 37°C, 0.1% CO2. All media were supplemented with antibiotics (1000 U/ml penicillin and 0.1 mg/ml streptomycin; Sigma-Aldrich). DharmaFECT (GE Healthcare Dharmacon, Lafayette, CO) and Lipofectamine (Life Technologies) were used to transfect siRNAs and plasmids, respectively.
3
Primary Culture of Gastric Mucosa
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Twenty-six ulcer patients who attended Gastroenterology Clinic of Mersin University Medical Faculty were included in this study. All ulcer patients gave written informed consent, and the project was reviewed and accepted by the Ethics Committee of Mersin University, Turkey. Gastroduodenal biopsies were obtained from patients undergoing upper endoscopy. Biopsy samples were taken either from the body of the stomach, or its antrum. The gastroduodenal biopsy samples were taken from grossly normal gastric mucosa. These endoscopic biopsy samples contained only mucosa (surface and deep glandular epithelium). The specimens were collected in Leibowitz's L-15 medium (Life Technologies) containing 1 % penicillin/streptomycin (Life Technologies) for transport of tissue biopsy sample from hospital to our laboratory. The primary culture was formed as described by Smooth DT from the ulcerous and control biopsy specimens (10) . Biopsy specimens were enzymatically disintegrated into Leibowitz L-15 solution consisting of type II collagenase (Biochrom 0.05 % and Dispase (120U). Then, within Ham's F12 medium (10 % fetal bovine serum, 1 % penicillin-streptomycin, 1 % Amphotericin-B), primary culture was incubated into four different 35-mm culture dishes. The culture was incubated at 37 o C in an incubator with 5 % CO 2 .
4
Live-cell Imaging of Cellular Dynamics
5
Imatinib Effects on SW480 Colon Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The chemical structure of imatinib (Sigma, with a purity ≥98%) is shown in Figure 1 . The colon cancer cell line SW480 was provided by the Shanghai Cell Bank of the Chinese Academy of Sciences. Fetal bovine serum (FBS) was purchased from Hyclone, Leibowitz’s L-15 medium and puromycin were bought from Gibco company, and rabbit anti-GAPDH, anti-HGF, anti-p21, and anti-p27 were purchased from Sigma-Aldrich.
6
Culturing Mammalian and Insect Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Baby hamster kidney cells stably expressing T7 polymerase (BHK/T7-9)32 (link) (kindly provided by Naoto Ito Gifu, Japan) were cultured in minimum essential medium supplemented with 10% fetal calf serum (FCS; Biowest, Nuaillé, France) and Tryptose phosphate broth. Vero E6 cells (kindly provided by Stephen Goodbourn, London, UK) were grown in Dulbecco's modified Eagle's medium (DMEM) (Gibco, Life Technologies, Cergy-Pontoise, France) supplemented with 10% FCS. Murine fibroblastic L929 cells and murine macrophage RAW 264.7 cells (ATCC, LGC Standards, Molsheim, France) were cultured in DMEM supplemented with L-glutamine and 10% FCS. Mammalian cells lines were maintained at 37 °C under 5% CO2 in medium supplemented with 10 IU of penicillin and 10 µg of streptomycin per mL. Aedes albopictus-derived C6/36 cells were grown in Leibowitz's L-15 medium (Gibco, Life Technologies, Cergy-Pontoise, France) supplemented with 10% FCS (Lonza, Verviers, Belgium) and 2% Tryptose phosphate broth (Sigma-Aldrich, Lyon, France) and incubated at 28 °C without CO2.
7
Live-cell Imaging of Cellular Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were imaged using a Nikon TE-2000E inverted microscope with a 603 1.49 NA TIRF objective, Andor Revolution XD spinning disk confocal system, and 488 and 568 nm solid-state lasers. Cells were imaged in Opti-MEM or Leibowitz's L15 medium (Gibco), 5% FBS, at 37°C. Time lapses were acquired using an Andor iXon+ EM-CCD camera using Andor IQ. Original 16-bit tif files acquired directly from the camera were used for image analysis.
8
Culturing Xenopus laevis Eye Primordia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Eye primordia were dissected from stage 24 X. laevis embryos and plated on 50 mm glass-bottom dishes (MatTek) coated with 10 mg ml−1 poly-l -lysine and 10 mg ml−1 laminin (both from Sigma). Explants were cultured at 20°C in 60% Leibowitz's L15 medium (Gibco) containing 5% penicillin/streptomycin/fungizone (Sigma cat. no. P4458) and 50 µg ml−1 gentamycin (GE Healthcare) for 16–18 h.
9
Embryonic Spinal Cord Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Our embryonic dissociation protocol was adapted from one previously used in the lab [39] (link). Embryos were grown to 20 hpf and anesthetized using Finquel (MS-222) (Argent Laboratories, Redmond, WA, USA). To obtain isolated spinal cords, embryos were incubated in 7.5× pancreatin (MP Biomedicals, Irvine, CA, USA) in zebrafish physiological saline [36] until tissues began to separate (about 1 minute). Embryos were then triturated with Pasteur pipettes of decreasing size. Isolated spinal cords were transferred to Leibowitz's L15 medium (Gibco/Life Technologies, Grand Island, NY, USA) containing 10% fetal bovine serum for 1–2 minutes to inactivate the pancreatin, after which they were stored in TriReagent (Molecular Research Center, Inc., Cincinnati, OH, USA) at −80°C.
10
Derivatization and Analysis of Steroids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fetal bovine serum (FBS), Leibowitz’s L-15 medium, the Roswell Park Memorial Institute (RPMI) 1640 medium, Ham’s F12 medium, Eagle’s minimal essential medium (EMEM), Dulbecco’s modified Eagle’s medium (DMEM), the Opti-MEM medium, a penicillin/streptomycin solution and trypsin were obtained from GIBCO (Grand Island, NY, USA). Trout serum was purchased from Caisson Laboratories (Smithfield, VA, USA). Mouse epidermal growth factor (EGF) and HEPES were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Sodium bicarbonate, bovine insulin, DHT, DHT-D3 solution, methyl-tertiary-butyl ether (MTBE), trimethylamine (TEA), tetrahydrofuran (THF), 2- PA, 4-(dimethylamino) pyridine (DMAP), 2-methyl-6-nitrobenzoic anhydride (MNBA), and acetic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA), and HPLC-grade formic acid was purchased from Fisher Scientific (Pittsburgh, PA, USA). MS-grade methanol and water were obtained from VWR (Westchester, NY, USA). The stock solution and internal standard were prepared in methanol. The derivatization reagent was prepared by dissolving 25.0 mg of PA, 10.0 mg of DMAP, and 20.0 mg of MNBA in 1 mL of THF (Yamashita et al., 2009) and vortexing. Then, the mixture was left at room temperature for at least 5 min before the sample pretreatment.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!