Iscove s modified dulbecco s media, by Thermo Fisher Scientific - Product details

1

Intracellular Cytokine Analysis of Blood and BAL Samples

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Directly after collection, blood and BAL samples were added to an equal volume (100 μL/tube) to IMDM (Iscove’s Modified Dulbecco’s Media, Thermo Fisher Scientific, Waltham, MA, USA) with or without 200 ng/mL of Escherichia coli LPS (E. coli K12 LPS, Invivogen, San Diego, CA, USA), then incubated for 4 h at 37 °C and 5% CO₂ [23 (link)]. Samples drawn to measure the intracellular cytokine synthesis of TNF-α were incubated with a Golgi inhibitor (GolgiStop™ 2 μM, Becton Dickinson, San Jose, CA, USA) for 4 h at 37 °C and 5% CO₂.

Bendib I., Beldi-Ferchiou A., Schlemmer F., Maitre B., Surenaud M., Hüe S., Carteaux G., Razazi K., Lelièvre J.D., Lévy Y., Mekontso Dessap A., Godot V, & de Prost N. (2021). Functional Ex Vivo Testing of Alveolar Monocytes in Patients with Pneumonia-Related ARDS. Cells, 10(12), 3546.

+ Open protocol

+ Expand

2

Culturing and Transfecting Jurkat T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

E6.1 Jurkat T cells were maintained at 37°C in Iscove’s modified Dulbecco’s media (Thermo Fisher Scientific) supplemented with 10% FBS (Sigma-Aldrich), sodium pyruvate, l-glutamine, penicillin-streptomycin, and MEM nonessential amino acids solution (Thermo Fisher Scientific). Transfections were performed with 1.0 × 10⁶ cells/ml and 2 to 3 µg plasmid DNA by nucleofection using Amaxa kit V (Lonza). Primary mouse T cells were obtained via dissection of mouse inguinal, axillary, brachial, and mesenteric lymph nodes. CD8⁺ T cells were isolated via negative selection using a CD8a⁺ T cell isolation kit (Kit II, MACS; Miltenyi Biotec) and grown in DMEM supplemented with 5% FBS, l-glutamine, and penicillin-streptomycin. The cells were then activated for 72 h with plate-bound 2.5 µg/ml anti-mouse CD3 and 1 µg/ml anti-mouse CD28 antibodies (BD), followed by stimulation with 50 U/ml recombinant interleukin-2 (R&D Systems) every 48 h.

Murugesan S., Hong J., Yi J., Li D., Beach J.R., Shao L., Meinhardt J., Madison G., Wu X., Betzig E, & Hammer J.A. (2016). Formin-generated actomyosin arcs propel T cell receptor microcluster movement at the immune synapse. The Journal of Cell Biology, 215(3), 383-399.

+ Open protocol

+ Expand

3

Isolation of Human Neutrophils

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fresh samples of peripheral blood from healthy volunteers was collected in 10 mL heparin or EDTA tubes (Research Blood Components LLC, Allston, MA). Protocols were approved by the institutional review board at Massachusetts General Hospital (MGH). Blood was utilized within 6 h after the blood draw, except for experiments with CGD samples that had both CGD and control blood shipped overnight from the National Institutes of Health (NIH). Neutrophils were isolated using the EasySep Direct Human Neutrophil Isolation Kit per the manufacturer’s protocol (STEMCELL Technologies). Isolated neutrophils were stained with Hoechst (ThermoFisher Scientific) and re-suspended in Iscove’s Modified Dulbecco’s Media with 20% fetal bovine serum (ThermoFisher Scientific).

Alex H., Scherer A., Kreuzburg S., Abers M.S., Zerbe C.S., Dinauer M.C., Mansour M.K, & Irimia D. (2020). Neutrophil swarming delays the growth of clusters of pathogenic fungi. Nature Communications, 11, 2031.

+ Open protocol

+ Expand

4

Culturing Human and Mouse Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human LSCs were cultured in StemSpan™ Serum-Free Expansion Medium II (STEMCELL, USA) with penicillin (100 U/mL), streptomycin (100 mg/mL), stem cell factor (SCF, 20 ng/ml), thrombopoietin (TPO, 20 ng/ml), Flt3-L (20 ng/ml), IL-3 (10 ng/ml), and IL-6 (10 ng/ml). Mouse LSCs were cultured in Iscove’s Modified Dulbecco’s Media (Thermo Fisher Scientific, USA) with 10% fetal bovine serum (FBS), penicillin (100 U/mL), streptomycin (100 mg/mL), SCF (20 ng/ml), TPO (20 ng/ml), Flt3-L (20 ng/ml), IL-3 (10 ng/ml), and IL-6 (10 ng/ml). Human and mouse ILC1s or NK cells were cultured in RPMI 1640 (Thermo Fisher Scientific, USA) with 10% FBS, penicillin (100 U/mL), streptomycin (100 mg/mL), IL-12 (10 ng/ml), and IL-15 (100 ng/ml). The mouse AML cell line C1498 (American Type Culture Collection) was cultured in RPMI 1640 with 10% FBS, penicillin (100 U/mL), and streptomycin (100 mg/mL). Cultures were incubated at 37°C in a humidified atmosphere of 5% CO2. All cytokines were from PeproTech. penicillin and streptomycin were from Thermo Fisher Scientific.

Li Z., Ma R., Ma S., Tian L., Lu T., Zhang J., Mundy-Bosse B.L., Zhang B., Marcucci G., Caligiuri M.A, & Yu J. (2022). ILC1s Control Leukemia Stem Cell Fate and Limit Development of AML. Nature immunology, 23(5), 718-730.

+ Open protocol

+ Expand

5

Culturing Human Lymphoma Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human lymphoma cell lines include cHL-derived (L1236, U-HO1, SUP-HD1, L591, L428, HDLM2, L540, and KM-H2), NLPHL-derived (DEV) and NHL/acute leukemia-derived cell lines (SU-DHL-6, SU-DHL-10, Toledo, U937, JeKo-1, NU-BL-1, DAUDI, Jurkat, and a pre-B acute lymphoblastic leukemia). All cells were cultured in RPMI-1640 (Corning cellgro ®, MA, USA) containing 10% to 20% fetal bovine serum and 100 μg/mL penicillin/streptomycin in 5% CO₂ incubator at 37 °C, with the exception of U-HO1 cells that were cultured in a 4:1 mixture of 80% Iscove’s Modified Dulbecco’s Media (Thermo Fisher Scientific Inc., Wilmington, MA, USA) and RPMI-1640 containing 20% FBS plus 2mM L-glutamine. SUP-HD1 cells were cultured in 80% McCoy’s 5A (Thermo Fisher) with 20% FBS. ABVD (doxorubicin, bleomycin, vinblastine, and dacarbazine) were purchased from Sigma Aldrich (St. Louis, MO, USA).

Li P.C., Siddiqi I.N., Mottok A., Loo E.Y., Wu C.H., Cozen W., Steidl C, & Shih J.C. (2017). Monoamine Oxidase A is Highly Expressed in Classical Hodgkin Lymphoma. The Journal of pathology, 243(2), 220-229.

+ Open protocol

+ Expand

6

Culturing Human and Mouse Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human LSCs were cultured in StemSpan™ Serum-Free Expansion Medium II (STEMCELL, USA) with penicillin (100 U/mL), streptomycin (100 mg/mL), stem cell factor (SCF, 20 ng/ml), thrombopoietin (TPO, 20 ng/ml), Flt3-L (20 ng/ml), IL-3 (10 ng/ml), and IL-6 (10 ng/ml). Mouse LSCs were cultured in Iscove’s Modified Dulbecco’s Media (Thermo Fisher Scientific, USA) with 10% fetal bovine serum (FBS), penicillin (100 U/mL), streptomycin (100 mg/mL), SCF (20 ng/ml), TPO (20 ng/ml), Flt3-L (20 ng/ml), IL-3 (10 ng/ml), and IL-6 (10 ng/ml). Human and mouse ILC1s or NK cells were cultured in RPMI 1640 (Thermo Fisher Scientific, USA) with 10% FBS, penicillin (100 U/mL), streptomycin (100 mg/mL), IL-12 (10 ng/ml), and IL-15 (100 ng/ml). The mouse AML cell line C1498 (American Type Culture Collection) was cultured in RPMI 1640 with 10% FBS, penicillin (100 U/mL), and streptomycin (100 mg/mL). Cultures were incubated at 37°C in a humidified atmosphere of 5% CO2. All cytokines were from PeproTech. penicillin and streptomycin were from Thermo Fisher Scientific.

Li Z., Ma R., Ma S., Tian L., Lu T., Zhang J., Mundy-Bosse B.L., Zhang B., Marcucci G., Caligiuri M.A, & Yu J. (2022). ILC1s Control Leukemia Stem Cell Fate and Limit Development of AML. Nature immunology, 23(5), 718-730.

+ Open protocol

+ Expand

7

Targeting Myeloma Cells with NM

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

MM1.S human MM 63 (link) and 5TGM1 murine MM 64 (link) cell lines, either naïve or carrying Click Beetle Red (CBR) luciferase and Green Fluorescent Protein (GFP) reporters (MM1.S/CBR/GFP and 5TGM/CBR/GFP, respectively), as well as MM1.S/CBR/GFP resistant to RaST and MM1.S/CBR/GFP without CD49d, were generously provided by Dr. DiPersio (Washington University School of Medicine, WUSM, St. Louis). Cells were routinely cultured in complete medium (CM) consisting of Iscove's Modified Dulbecco's Media (Thermo Fisher Scientific, Waltham, MA) supplemented with 10% heat-inactivated fetal bovine serum (FBS, ThermoFisher Scientific, Waltham, MA) and 50 µg/mL Gentamycin (Thermo Fisher Scientific, Waltham, MA). The cells were routinely washed in phosphate-buffered saline (PBS, Thermo Fisher Scientific, Waltham, MA), pH 7.4. VLA-4 and αvß3 targeted NM were loaded with TC (Bis(cyclopentadienyl)titanium(IV) dichloride, Sigma-Aldrich, St. Louis, MO) 43 , 46 (link). Protein content was routinely measured with Pierce^TM bicinchoninic acid (BCA) assay (Thermo Fisher Scientific Waltham, MA).

Zheleznyak A., Mixdorf M., Marsala L., Prior J., Yang X., Cui G., Xu B., Fletcher S., Fontana F., Lanza G, & Achilefu S. (2021). Orthogonal targeting of osteoclasts and myeloma cells for radionuclide stimulated dynamic therapy induces multidimensional cell death pathways. Theranostics, 11(16), 7735-7754.

+ Open protocol

+ Expand

8

Reconstitution of Cell-Adhesion Molecules

Check if the same lab product or an alternative is used in the 5 most similar protocols

1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] (nickel salt) (DOGS-NTA (Ni²⁺)) and 1,2-dioleoyl-sn-glycero-phospho-ethanolamine-3-N-(cap biotinyl) (biotin-DOPE) were purchased from Avanti Polar Lipids (Alabaster, USA), and neutravidin from Life Technologies. Recombinant stromal cell-derived factor-1α (SDF1α) with and without biotin tags and human N-cadherin with histidine tag were purchased from Almac Group (Craigavon, UK) and R&D Systems Inc. (Wiesbaden, Germany), respectively. For all cell experiments, Iscove's Modified Dulbecco's Media from Life Technologies was used.

Burk A.S., Monzel C., Yoshikawa H.Y., Wuchter P., Saffrich R., Eckstein V., Tanaka M, & Ho A.D. (2015). Quantifying Adhesion Mechanisms and Dynamics of Human Hematopoietic Stem and Progenitor Cells. Scientific Reports, 5, 9370.

+ Open protocol

+ Expand

9

Functionalized Lipid Membrane Preparation

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] (nickel salt) (DOGS-NTA (Ni²⁺)) and 1,2-dioleoyl-sn-glycero-phospho-ethanolamine-3-N-(cap biotinyl) (biotin-cap-DOPE) were purchased from Avanti Polar Lipids (Alabaster, USA), and neutravidin from Life Technologies. Recombinant stromal cell-derived factor-1 (SDF1α) with and without biotin tags and human N-cadherin with histidin tag were purchased from Almac Group (Craigavon, UK) and R&D Systems Inc. (Wiesbaden, Germany), respectively. Plerixafor was purchased from Sigma and NOX-A12 was provided by NOXXON Pharma AG (Berlin, Germany) and used without further purification. The nucleotide sequence of NOX-A12 (5′-GCGUGGUGUGAUCUAGAUGUAUUGGCUGAUCCUAGUCAGGUACGC-3′) was obtained from in vitro selection experiments as described before^{51 (link)}. For all cell experiments, Iscove’s Modified Dulbecco’s Media from Life Technologies (Darmstadt, Germanz) was used.

Monzel C., Becker A.S., Saffrich R., Wuchter P., Eckstein V., Ho A.D, & Tanaka M. (2018). Dynamic cellular phenotyping defines specific mobilization mechanisms of human hematopoietic stem and progenitor cells induced by SDF1α versus synthetic agents. Scientific Reports, 8, 1841.

+ Open protocol

+ Expand

10

Gastric Cancer Cell Line Maintenance and Transfection

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human GC cell lines (SNU-5, SNU-16, and NCI-N87) and a normal gastric mucosa cell
line (HS 738 ST/Int) were purchased from American Type Culture Collection. Human
GC cell lines (MGC80-3 and SGC-7901) were provided by the Chinese Academy of
Sciences. HS 738 ST/Int was maintained in Dulbecco’s Modified Eagle Medium
(Invitrogen; Thermo Fisher Scientific, Inc). SNU-5 was cultured in Iscove’s
Modified Dulbecco’s Media (Invitrogen; Thermo Fisher Scientific, Inc), and
others were cultured in RPMI-1640 medium (Invitrogen; Thermo Fisher Scientific,
Inc) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Inc),
100 U/mL penicillin, and 100 μg/mL streptomycin (Solarbio). All cell lines were
placed in a humidified incubator with 5% CO₂ at 37 °C.
MicroRNA-12129 mimics, small interfering RNA-SIRT1, and negative control (NC)
mimics were purchased from GenePharma. The detailed sequences are described in
Table 1. Cell
lines (SNU-5, NCI-N87) were transfected in 6-well plates with the Lipofectamine
2000TM reagent (Invitrogen; Thermo Fisher Scientific, Inc) in accordance with
the manufacturer protocol.

Zhang W., Liao K, & Liu D. (2020). MiRNA-12129 Suppresses Cell Proliferation and Block Cell Cycle Progression by Targeting SIRT1 in GASTRIC Cancer. Technology in Cancer Research & Treatment, 19, 1533033820928144.

+ Open protocol

+ Expand

Iscove s modified dulbecco s media

Lab products found in correlation

Close spelling variants of this product

24 protocols using iscove s modified dulbecco s media

Intracellular Cytokine Analysis of Blood and BAL Samples

Culturing and Transfecting Jurkat T Cells

Isolation of Human Neutrophils

Culturing Human and Mouse Immune Cells

Culturing Human Lymphoma Cell Lines

Culturing Human and Mouse Immune Cells

Targeting Myeloma Cells with NM

Reconstitution of Cell-Adhesion Molecules

Functionalized Lipid Membrane Preparation

Gastric Cancer Cell Line Maintenance and Transfection

About PubCompare

Ready to get started?