DMEM high glucose (4.5 g/L) with glutamine (BE12-604 F), HEPES pH 7.4 (BE17-737E) and Penicillin/Streptomycin (DE17-602E) were purchased from Lonza (Cologne, Germany). Fetal Bovine Serum (FBS HyClone, SH30070.03) was obtained from GE Healthcare Europe (Vienna, Austria), the cell viability assay Cell Counting Kit-8 (CCK-8) was from Dojindo Molecular Technologies Japan (CK04, Dojindo EU, Munich, Germany). Cell culture expendables were purchased from Falcon (Corning, Germany). Glutamic acid was purchased from Sigma (G5513, Vienna, Austria), LMP (Neurocil®) was purchased from Bayer Pharma (Leverkusen, Germany).
Be17 737e
Manufactured by Lonza
Sourced in Germany
The BE17-737E is a laboratory equipment product manufactured by Lonza. It is designed for specific lab-based applications. No further details about its core function or intended use can be provided in an unbiased and factual manner without potential for extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Moflo astrio, by Beckman Coulter (2 mentions)
Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (2 mentions)
Glutamic acid, by Merck Group (1 mentions)
Be12 604f, by Lonza (1 mentions)
Hyclone fetal bovine serum fbs, by GE Healthcare (1 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
Doxycycline, by Merck Group (1 mentions)
Chir 124, by Selleck Chemicals (1 mentions)
Pf 477736, by Selleck Chemicals (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
Penicillin, by Lonza (1 mentions)
Streptomycin, by Lonza (1 mentions)
Sodium pyruvate, by Lonza (1 mentions)
Be17 605e, by Lonza (1 mentions)
De17 602e, by Lonza (1 mentions)
Be12 125f, by Lonza (1 mentions)
Xtt assay, by Roche (1 mentions)
Ly6c apc clone hk1, by BioLegend (1 mentions)
5 protocols using be17 737e
1
Cell Viability Assay Protocol
2
Cell Culture Conditions for Hematological and Embryonic Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were cultured at 37 °C in a humidified atmosphere containing 5% CO2, except for mouse pre-B cells (7.5% CO2). Human Burkitt lymphoma, Nalm6 cells and murine pre-B cell lines were cultured in RPMI-1640 medium (Sigma-Aldrich, R0883) supplemented with 10% FCS (Gibco, 10270-106), 2 mM l -glutamine (Sigma, G7513), 100 U/ml penicillin and 100 μg/ml streptomycin (Sigma, P0781) and 50 μM 2-mercaptoethanol (Sigma, M3148). For murine pre-B cells and primary bone marrow culture, 1 mM sodium pyruvate (Gibco, 11360-039), 1× non-essential amino acids (Gibco, 11140-35) and IL-7 (supernatant of IL-7 expressing J558L cells) was added in addition. Primary FACS-sorted murine pro B cells were cultured in DMEM medium (Sigma, D5671), supplemented with 10% FCS, 100 U/ml penicillin and 100 μg/ml streptomycin, 2 mM l ‐glutamine, 10 mM Hepes (LONZA, BE17-737E), 1 mM sodium pyruvate, 1× non-essential amino acids and 50 μM 2-mercaptoethanol. Human embryonic kidney cells (HEK293T) were grown in DMEM medium, supplemented with 10% FCS, 100 U/ml penicillin and 100 μg/ml streptomycin and 2 mM l ‐glutamine. Reagents: PF-477736 (Selleckchem S2904), CHIR-124 (Selleckchem S2683), QVD (SML0063, Sigma), DMSO (D5879, Sigma), Doxycycline (D9891, Sigma), anti-mouse CD40 (102908, biolegend, HM40-3).
3
Cytotoxicity Evaluation of NovioSense Glucose Sensor
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytotoxicity tests were outsourced and conducted
by Toxikon Europe
NV, Belgium according to ISO 10993-5, 2009, Biological Evaluation
of Medical Devices; Part 5: Tests in vitro cytotoxicity and ISO 10993-12,
2012, Biological Evaluation of Medical Devices; Part 12: Sample Preparation
and Reference Materials. The study was conducted in accordance with
Good Laboratory Practice (GLP). The test was based on the measurement of
viability of L929 mouse fibroblasts (105 cells/mL) in response
to an extract of the NovioSense Glucose Sensor functionalized with
GOx dummy device via XTT assay (Roche). The NovioSense Glucose Sensor
devices were extracted in 0.9% sodium chloride at a ratio of 60 cm2/20 mL. The NovioSense Glucose Sensor device extract was finally
diluted in serum-supplemented Minimum Essential Medium (MEM-complete)
at a ratio of 1/4. The L929 cells were exposed in quadruplicate at
this 1/4 dilution. MEM-complete contained following: MEM (LONZA:BE12-125F),
penicillin/streptomycin (LONZA: DE17-602E), fetal bovine serum (Greiner-bio-one
FBSEU500),l -glutamine (LONZA:BE17-605E), Hepes buffer (LONZA:
BE17–737E).
by Toxikon Europe
NV, Belgium according to ISO 10993-5, 2009, Biological Evaluation
of Medical Devices; Part 5: Tests in vitro cytotoxicity and ISO 10993-12,
2012, Biological Evaluation of Medical Devices; Part 12: Sample Preparation
and Reference Materials. The study was conducted in accordance with
Good Laboratory Practice (GLP). The test was based on the measurement of
viability of L929 mouse fibroblasts (105 cells/mL) in response
to an extract of the NovioSense Glucose Sensor functionalized with
GOx dummy device via XTT assay (Roche). The NovioSense Glucose Sensor
devices were extracted in 0.9% sodium chloride at a ratio of 60 cm2/20 mL. The NovioSense Glucose Sensor device extract was finally
diluted in serum-supplemented Minimum Essential Medium (MEM-complete)
at a ratio of 1/4. The L929 cells were exposed in quadruplicate at
this 1/4 dilution. MEM-complete contained following: MEM (LONZA:BE12-125F),
penicillin/streptomycin (LONZA: DE17-602E), fetal bovine serum (Greiner-bio-one
FBSEU500),
BE17–737E).
4
Isolation and Purification of Microglia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Microglia were isolated as previously described [54 ]. After perfusion, brains were removed from the skull and kept in cold medium A (HBSS (Gibco, 14170-088) with 0.6% glucose (Sigma, G8769) and 7.5 mM HEPES (Lonza, BE17-737E)). All subsequent steps were performed on ice, centrifugation was at 4 °C. Brains were dissociated using a Potter–Elvehjem tissue homogenizer after which the homogenate was passed over a 70 µM cell strainer (Corning, 352350) and pelleted by centrifugation at 220×g for 10 min. Next, myelin was removed by resuspending the pellet in 25 mL 24% Percoll (Fisher, 17-0891-01) in medium A (1 × final concentration) with 3 mL PBS layered on top, followed by centrifugation for 20 min at 950×g (acceleration 4 and brake 0). The microglia enriched cell pellets were incubated with CD11b-PE (clone M1/70, eBiosciences, 12-0112-82), CD45-FITC (clone 30-F11, eBiosciences, 11-0451-82), and Ly6c-APC (clone HK1.4, Biolegend, 128016) antibodies for 30 min on ice. Then the cells were washed once in medium A without phenol red and filtered into FACS tubes. Microglia were sorted by gating the DAPInegCD11bhighCD45intLy6cneg cells using the Beckman Coulter MoFlo Astrios or XDP. Microglia were collected in siliconized Eppendorf tubes (Sigma, T3406-250EA) containing medium A. Flow cytometry data were analyzed using FlowJo Analysis Software.
5
Microglia Isolation from CNS Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Microglia were isolated from whole brain (LPS) or spinal cord (EAE) as previously described in detail [21 (link)]. The whole isolation procedure was performed on ice. Mice were perfused with PBS (Lonza, BE17-512F) and CNS tissue was mechanically dissociated in HBSS (Gibco, 14,170–088) containing 0.6% glucose (Sigma-Aldrich, G8769) and 15 mM HEPES (Lonza, BE17-737E). Myelin was removed by 24.4% percoll (GE Healthcare, 17-0891-01) density gradient centrifugation at 950 g for 20 min at 4 °C. Fc receptors were blocked with anti-CD16/32 (5 μg/ml, clone 93, eBioscience, 14-0161-85), and cells were stained with anti-CD11b-APC-Cy7 (1 μg/ml, clone M1/70, eBioscience, A15390), anti-CD45-PE-Cy7 (1 μg/ml, clone 30-F11, eBioscience, 25-0451-82), anti-Ly6C-APC (1.5 μg/ml, clone HK1.4, Biolegend, 128,016), and anti-VISTA-PE (20 μg/ml, clone MIH63, Biolegend, 150,204) antibodies 30 min at 4 °C in HBSS without phenol red (Gibco, 14,175-053) containing 0.6% glucose, 15 mM HEPES, and 1 mM EDTA (Invitrogen, 15,575-020). Microglia were sorted on a MoFlo Astrios (Beckman Coulter) in siliconized tubes containing RNAlater (Qiagen, 76,104), centrifuged at 5000 g, and lysed in RLT + lysis buffer (Qiagen, 74,034).
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!